Recombinant swinepox virus

ABSTRACT

The present invention relates to a recombinant swinepox virus capable of replication comprising foreign DNA inserted into a site in the swinepox viral DNA which is not essential for replication of the swinepox virus. The invention further relates to homology vectors which produce recombinant swinepox viruses by inserting foreign DNA into swinepox viral DNA

This application is a U.S. national stage application under 35 USC 371of international PCT/US94/08277, filed Jul. 22, 1994, which is acontinuation-in-part of U.S. Ser. No. 07/820,154 filed Jan. 13, 1992,U.S. Pat. No. 5,382,425, and U.S. Ser. No. 08/097,554, filed Jul. 22,1993, U.S. Pat. No. 5,869,312, the contents of which are incorporated byreference into the present application. Within this application severalpublications are referenced by arabic numerals within parentheses. Fullcitations for these publications may be found at the end of thespecification immediately preceding the claims. The disclosures of thesepublications are hereby incorporated by reference into this applicationin order to more fully describe the state of the art to which thisinvention pertains.

BACKGROUND OF THE INVENTION

Swinepox virus (SPV) belongs to the family Poxviridae. Viruses belongingto this group are large, double-stranded DNA viruses thatcharacteristically develop in the cytoplasm of the host cell. SPV is theonly member of the genus Suipoxvirus. Several features distinguish SPVfrom other poxviruses. SPV exhibits species specificity (18) compared toother poxviruses such as vaccinia which exhibit a broad host range. SPVinfection of tissue culture cell lines also differs dramatically fromother poxviruses (24). It has also been demonstrated that SPV does notexhibit antigenic cross-reactivity with vaccinia virus and shows nogross detectable homology at the DNA level with the ortho, lepori, avior entomopox virus groups (24). Accordingly, what is known and describedin the prior art regarding other poxviruses does not pertain a priori toswinepox virus.

SPV is only mildly pathogenic, being characterized by a self-limitinginfection with lesions detected only in the skin and regional lymphnodes. Although the SPV infection is quite limited, pigs which haverecovered from SPV are refractory to challenge with SPV, indicatingdevelopment of active immunity (18).

The present invention concerns the use of SPV as a vector for thedelivery of vaccine antigens and therapeutic agents to swine. Thefollowing properties of SPV support this rationale: SPV is only mildlypathogenic in swine, SPV is species specific, and SPV elicits aprotective immune response. Accordingly, SPV is an excellent candidatefor a viral vector delivery system, having little intrinsic risk whichmust be balanced against the benefit contributed by the vector's vaccineand therapeutic properties.

The prior art for this invention stems first from the ability to cloneand analyze DNA while in bacterial plasmids. The techniques that areavailable are detailed for the most part in Maniatis et al., 1983 andSambrook et al., 1989. These publications teach state of the art generalrecombinant DNA techniques.

Among the poxviruses, five (vaccinia, fowlpox, canarypox, pigeon, andraccoon pox) have been engineered, previous to this disclosure, tocontain foreign DNA sequences. Vaccinia virus has been used extensivelyto vector foreign genes (25) and is the subject of U.S. Pat. Nos.4,603,112 and 4,722,848. Similarly, fowlpox has been used to vectorforeign genes and is the subject of several patent applications EPA 0284 416, PCT WO 89/03429, and PCT WO 89/12684. Raccoon pox (10) andCanarypox (31) have been utilized to express antigens from the rabiesvirus. These examples of insertions of foreign genes into poxviruses donot include an example from the genus Suipoxvirus. Thus, they do notteach methods to genetically engineer swinepox viruses, that is, whereto make insertions and how to get expression in swinepox virus.

The idea of using live viruses as delivery systems for antigens has avery long history going back to the first live virus vaccines. Theantigens delivered were not foreign but were naturally expressed by thelive virus in the vaccines. The use of viruses to deliver foreignantigens in the modern sense became obvious with the recombinantvaccinia virus studies. The vaccinia virus was the vector and variousantigens from other disease causing viruses were the foreign antigens,and the vaccine was created by genetic engineering. While the conceptbecame obvious with these disclosures, what was not obvious was theanswer to a more practical question of what makes the best candidatevirus vector. In answering this question, details of the pathogenicityof the virus, its site of replication, the kind of immune response itelicits, the potential it has to express foreign antigens, itssuitability for genetic engineering, its probability of being licensedby regulatory agencies, etc, are all factors in the selection. The priorart does not teach these questions of utility.

The prior art relating to the use of poxviruses to deliver therapeuticagents relates to the use of a vaccinia virus to deliver interleukin-2(12). In this case, although the interleukin-2 had an attenuating effecton the vaccinia vector, the host did not demonstrate any therapeuticbenefit.

The therapeutic agent that is delivered by a viral vector of the presentinvention must be a biological molecule that is a by-product of swinepoxvirus replication. This limits the therapeutic agent in the firstanalysis to either DNA, RNA or protein. There are examples oftherapeutic agents from each of these classes of compounds in the formof anti-sense DNA, anti-sense RNA (16), ribozymes (34), suppressor tRNAs(2), interferon-inducing double stranded RNA and numerous examples ofprotein therapeutics, from hormones, e.g., insulin, to lymphokines,e.g., interferons and interleukins, to natural opiates. The discovery ofthese therapeutic agents and the elucidation of their structure andfunction does not make obvious the ability to use them in a viral vectordelivery system.

SUMMARY OF THE INVENTION

The invention provides a recombinant swinepox virus capable ofreplication which comprises swinepox viral DNA and foreign DNA encodingRNA which does not naturally occur in an animal into which therecombinant swinepox virus is introduced. The foreign DNA is insertedinto the swinepox viral DNA at a site which is not essential forreplication of the swinepox virus and is under the control of apromoter.

This invention provides a homology vector for producing a recombinantswinepox virus by inserting foreign DNA into the genomic DNA of aswinepox virus which comprises a double-stranded DNA molecule. Thismolecule consists essentially of double-stranded foreign DNA encodingRNA which does not naturally occur in an animal into which therecombinant swinepox virus is introduced. At one end of this foreign DNAis double-stranded swinepox viral DNA homologous to genomic DNA locatedat one side of a site on the genomic DNA which is not essential forreplication of the swinepox virus. At the other end of the foreign DNAis double-stranded swinepox viral DNA homologous to genomic DNA locatedat the other side of the same site on the genomic DNA.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A and 1B show a detailed diagram of SPV genomic DNA (Kaszastrain) including the unique long and Terminal repeat (TR) regions. Arestriction map for the enzyme HindIII is indicated (23). Fragments arelettered in order of decreasing size. Note that the terminal repeats aregreater than 2.1 kb but less than 9.7 kb in size.

FIGS. 2A and 2B show the DNA sequence from homology vector 515-85.1. Thesequence of two regions of the homology vector 515-85.1 are shown. Thefirst region (FIG. 2A) (SEQ ID NO:1) covers a 599 base pair sequencewhich flanks the unique AccI site as indicated in FIG. 3. The beginning(Met) and end (Val) of a 115 amino acid ORF is indicated by thetranslation of amino acids below the DNA sequence. The second region(FIG. 2B) (SEQ ID NO:3) covers the 899 base pairs upstream of the uniqueHindIII site as indicated in FIG. 3. The beginning (Asp) and end (Ile)of a 220 amino acid ORF is indicated by the translation of amino acidsbelow the DNA sequence.

FIGS. 3A, 3B, and 3C show the homology which exists between the 515.85.1ORF and the Vaccinia virus 01L ORF. FIG. 3A shows two maps: The firstline of FIG. 3A is a restriction map of the SPV HindIII M fragment andthe second is a restriction map of the DNA insertion in plasmid515-85.1. The location of the 515-85.1 [VV 01L-like] ORF is alsoindicated on the map. The locations of the DNA sequences shown in FIGS.3B and 3C are indicated below the map by heavy bars in FIG. 3A. FIG. 3Bshows the homology between the VV 01L ORF (SEQ ID NO:5) and the 515-85.1ORF (SEQ ID NO:6) at their respective N-termini. FIG. 3C shows thehomology between the VV 01L ORF (SEQ ID NO:7) and the 515-85.1 ORF (SEQID NO:8) at their respective C-termini.

FIGS. 4A, 4B, and 4D show a description of the DNA insertion in HomologyVector 520-17.5. FIG. 4A contains a diagram showing the orientation ofDNA fragments assembled in plasmid 520-17.5 and table indicating theorigin of each fragment. FIG. 4B shows the sequences located at each ofthe junctions A and B between fragments, and FIG. 4C shows the sequenceslocated at Junctions C and D (SEQ ID NO's: 9, 10, 13, and 16). FIGS. 4Band 4C further describe the restriction sites used to generate eachfragment as well as the synthetic linker sequences which were used tojoin the fragments are described for each junction. The synthetic linkersequences are underlined by a heavy bar. The location of several genecoding regions and regulatory elements are also given. The following twoconventions are used: numbers in parenthesis ( ) refer to amino acids,and restriction sites in brackets [ ] indicate the remnants of siteswhich were destroyed during construction. The following abbreviationsare used, swinepox virus (SPV), early promoter 1 (EPI), late promoter 2(LP2), lactose operon Z gene (lacZ), and Escherichia coli (E. coli).

FIGS. 5A, 5B, 5C, and 5D shows a detailed description of the DNAinsertion in Homology Vector 538-46.16. FIG. 5A contains a diagramshowing the orientation of DNA fragments assembled in plasmid 538-46.16and a table indicating the origin of each fragment. FIG. 5B shows thesequences located at Junctions A and B between fragments, FIG. 5C showssequences located at Junction C and FIG. 5D shows sequences located atJunctions D and E (SEQ ID NO's: 17, 18, 21, 26, and 28). FIGS. 5B to 5Dalso describe the restriction sites used to generate each fragment aswell as the synthetic linker sequences which were used to join thefragments are described for each junction. The synthetic linkersequences are underlined by a heavy bar. The location of several genecoding regions and regulatory elements is also given. The following twoconventions are used: numbers in parenthesis ( ) refer to amino acids,and restriction sites in brackets [ ] indicate the remnants of siteswhich were destroyed during construction. The following abbreviationsare used, swinepox virus (SPV), pseudorabies virus (PRV), g50 (gpD),glycoprotein 63 (gp63), early promoter 1 (EP1), late promoter 1 (LP1)(SEQ ID NO: 46), late promoter 2 (LP2), lactose operon Z gene (lacZ),and Escherichia coli (E. coli).

FIG. 6

Western blot of lysates from recombinant SPV infected cells withanti-serum to PRV. Lanes (A) uninfected Vero cell lysate, (B) S-PRV-000(pseudorabies virus S62/26) infected cell lysate, (C) pre-stainedmolecular weight markers, (D) uninfected EMSK cell lysate, (E) S-SPV-000infected cell lysate, (F) S-SPV-003 infected cell lysate, (G) S-SPV-008infected cell lysate. Cell lysates were prepared as described in thePREPARATION OF INFECTED CELL LYSATES. Approximately 1/5 of the totallysate sample was loaded in each lane. FIG. 7

DNA sequence of NDV Hemagglutinin-Neuraminidase gene (HN) (SEQ ID NO:29). The sequence of 1907 base pairs of the NDV HN cDNA clone are shown.The translational start and stop of the HN gene is indicated by theamino acid translation below the DNA sequence.

FIGS. 8A, 8B, 8C, and 8D show a detailed description of the DNAinsertion in Homology Vector 538-46.26. FIG. 5A contains a diagramshowing the orientation of DNA fragments assembled in plasmid 538-46.26and table indicating the origin of each fragment. FIG. 8B shows thesequences located at Junctions A and B between fragments; FIG. 8C showsthe sequences located at Junctions C and D, FIG. 8D shows the sequenceslocated at Junction E (SEQ ID NO's: 31, 32, 34, 37, and 40). Therestriction sites used to generate each fragment as well as thesynthetic linker sequences which were used to join the fragments aredescribed for each junction in FIGS. 8B and 8D. The synthetic linkersequences are underlined by a heavy bar. The location of several genecoding regions and regulatory elements is also given. The following twoconventions are used: numbers in parenthesis ( ) refer to amino acids,and restriction sites in brackets [ ] indicate the remnants of siteswhich were destroyed during construction. The following abbreviationsare used, swinepox virus (SPV), Newcastle Disease virus (NDV),hemagglutinin-neuraminidase (HN), early promoter 1 (EP1), late promoter1 (LP1), late promoter 2 (LP2), lactose operon Z gene (lacZ), andEscherichia coli (E. coli).

FIGS. 9A, 9B, and 9C show a detailed description of Swinepox VirusS-SPV-010 and the DNA insertion in Homology Vector 561-36.26. FIG. 9Acontains a diagram showing the orientation of DNA fragments assembled inplasmid 561-36.26 and a table indicating the origin of each fragment.FIG. 9B shows the sequences located at Junctions A and B betweenfragments and FIG. 9C show the sequences located at junction C and D(SEQ ID. NO: 47, 48, 49,50). The restriction sites used to generate eachfragment as well as synthetic linker sequences which are used to jointhe fragments are described for each junction in FIGS. 9B and 9C. Thelocation of several gene coding regions and regulatory elements is alsogiven. The following two conventions are used: numbers in parentheses, (), refer to amino acids, and restriction sites in brackets, [ ],indicate the remnants of sites which are destroyed during construction.The following abbreviations are used: swinepox virus (SPV), Escherichiacoli (E. coli), thymidine kinase (TK), pox synthetic late promoter 1(LP1), base pairs (BP).

FIGS. 10A, 10B, 10C, and 10D show a detailed description of SwinepoxVirus S-SPV-011 and the DNA insertion in Homology Vector 570-91.21. FIG.10A contains a diagram showing the orientation of DNA fragmentsassembled in plasmid 570-91.21 and a table indicating the origin of eachfragment. FIG. 10B show the sequences located at Junctions A and Bbetween fragments; FIG. 10C shows the sequences located at Junction C,and FIG. 10D shows the sequences located at Junctions 10D and 10E (SEQID NO: 51, 52, 53, 54, 55). The restriction sites used to generate eachfragment as well as synthetic linker sequences which are used to jointhe fragments are described for each junction in FIGS. 10B to 10D. Thelocation of several gene coding regions and regulatory elements is alsogiven. The following two conventions are used: numbers in parentheses, (), refer to amino acids, and restriction sites in brackets, [ ],indicate the remnants of sites which are destroyed during construction.The following abbreviations are used: swinepox virus (SPV), pseudorabiesvirus (PRV), Escherichia coli (E. coli), pox synthetic late promoter 1(LP1), pox synthetic early promoter 2 (EP2) (SEQ ID NO: 45), gIII (gpC),base pairs (BP).

FIGS. 11, 11B, 11C and 11D show a detailed description of Swinepox VirusS-SPV-012 and the DNA insertion in Homology Vector 570-91.41. FIG. 11Acontains a diagram showing the orientation of DNA fragments assembled inplasmid 570-91.41 and a table indicating the origin of each fragment.FIG. 11B shows the sequences located at Junctions A and B betweenfragements, FIG. 11C shows the sequences located at Junction C, and FIG.11D shows the sequence located at Junctions D and E. (SEQ ID NO: 56, 57,58, 59, 60). The restriction sites used to generate each fragment aswell as synthetic linker sequences which are used to join the fragmentsare described for each junction in FIGS 11B to 11D. The location ofseveral gene coding regions and regulatory elements is also given. Thefollowing two conventions are used: numbers in parentheses, ( ), referto amino acids, and restriction sites brackets, [ ], indicate theremnants of sites which are destroyed during construction. The followingabbreviations are used: swinepox virus (SPV), pseudorabies virus (PRV),Escherichia coli (E. coli), pox synthetic late promoter 1 (LP1), poxsynthetic early promoter 1 late promoter 2 (EP1LP2) (SEQ ID NO: 43),gIII (gpC), base pairs (BP).

FIGS. 12, 12B, 12C and 12D show a detailed description of Swinepox VirusS-PRV-013 and the DNA insertion in Homology Vector 570-91.64. FIG. 12Acontains a diagram showing the orientation of DNA fragments assembled inplasmid 570-91.64 and a table indicating the origin of each fragment.FIG. 12B shows the sequences located at Junctions A and B betweenfragments, FIG. 12C shows the sequences located at Junction C, and FIG.12D shows the sequences located at Junctions D and E (SEQ ID NO: 61, 62,63, 64, 65). The restriction sites used to generate each fragment aswell as synthetic linker sequences which are used to join the fragmentsare described for each junction in FIGS. 12B to 12D. The location ofseveral gene coding regions and regulatory elements is also given. Thefollowing two conventions are used: numbers in parentheses, ( ), referto amino acids, and restriction sites in brackets, [ ], indicate theremnants of sites which are destroyed during construction. The followingabbreviations are used: swinepox virus (SPV), pseudorabies virus (PRV),Escherichia coli (E. coli), pox synthetic late promoter 1 (LP1), poxsynthetic late promoter 2 early promoter 2 (LP2EP2) (SEQ ID NO: 44),gIII (gpC) base pairs (BP).

FIGS. 13A, 13B, 13C and 13D show a detailed description of SwinepoxVirus S-PRV-014 and the DNA insertion in Homology Vector 599-65.25. FIG.13A contains a diagram showing the orientation of DNA fragmentsassembled in plasmid 599-65.25 and a table indicating the origin of eachfragment. FIG. 13B shows sequences located at Junctions A and B betweenthe fragments, FIG. 13C shows sequences located at Junction C, and FIG.13D shows sequences located at Junctions D and E. (SEQ ID NO: 66, 67,68, 69, 70). The restriction sites used to generate each fragment aswell as synthetic linker sequences which are used to join the fragmentsare described for each junction in FIGS. 13B to 13D. The location ofseveral gene coding regions and regulatory elements is also given. Thefollowing two conventions are used: numbers in parentheses, ( ), referto amino acids, and restriction sites in brackets, [ ], indicate theremnants of sites which are destroyed during construction. The followingabbreviations are used:

swinepox virus (SPV), infectious laryngotracheitis virus (ILT),Escherichia coli (E. coli), pox synthetic late promoter 1 (LP1), poxsynthetic early promoter 1 late promoter 2 (EP1LP2), glycoprotein G(gpG), polymerase chain reaction (PCR), base pairs (BP).

FIGS. 14A, 14B, 14C, and 14D show a derailed description of SwinepoxVirus S-SPV-016 and the DNA insertion in Homology Vector 624-20.1C. FIG.14A contains a diagram showing the orientation of DNA fragmentsassembled in plasmid 624-20.1C and a table indicating the origin of eachfragment. FIG. 14B shows the sequences located at Junctions A and Bbetweeen fragments; FIG. 14C shows the sequences located at Junction C,and FIG. 14D shows the sequences at Junctions D and E. (SEQ ID NO: 71,72, 73, 74, 75). The restriction sites are used to generate eachfragment as well as synthetic linker sequences which are used to jointhe fragments are described for each junction in FIGS. 14B to 14D. Thelocation of several gene coding regions and regulatory elements is alsogiven. The following two conventions are used: numbers in parentheses, (), refer to amino acids, and restriction sites in brackets, [ ],indicate the remnants of sites which are destroyed during construction.The following abbreviations are used: swinepox virus (SPV), infectiouslaryngotracheitis virus (ILT), Escherichia coli (E. coli), pox syntheticlate promoter 1 (LP1), pox synthetic late promoter 2 early promoter 2(LP2EP2), glycoprotein I (gpI), polymerase chain reaction (PCR), basepairs (BP).

FIGS. 15A, 15B, 15C and 15D show a detailed description of SwinepoxVirus S-SPV-017 and the DNA insertion in Homology Vector 614-83.18. FIG.15A contains a diagram showing the orientation of DNA fragmentsassembled in plasmid 614-83.18 and a table showing the origin of eachfragment. FIG. 15B shows the sequences located at Junctions A and Bbetween fragments, FIG. 15C shows the sequences at Junction C, and FIG.15D shows the sequences located at Junctions D and E. The restrictionsites used tc generate each fragment as well as synthetic linkersequences which are used to join the fragments are described for eachjunction in FIGS. 15B to 15D. The location of several gene codingregions and regulatory elements is also given. The following twoconventions are used: numbers in parentheses, ( ), refer to amino acids,and restriction sites in brackets, [ ], indicate the remnants of siteswhich are destroyed during construction. The following abbreviations areused: swinepox virus (SPV), infectious bovine rhinotracheitis virus(IBR), Escherichia coli (E. coli), pox synthetic late promoter 1 (LP1),pox synthetic late promoter 2 early promoter 2 (LP2EP2), glycoprotein G(gpG), polymerase chain reaction (PCR), base pairs (BP).

FIG. 16

Western blot of lysates from recombinant SPV infected cells withpolyclonal goat anti-PRV gIII (gpC). Lanes (A) S-PRV-002 (U.S. Pat. No.4,877,737, issued Oct. 31, 1989) infected cell lysate, (B) molecularweight markers, (C) mock-infected EMSK cell lysate, (D) S-SPV-003infected cell lysate, (E) S-SPV-008 infected cell lysate, (F) S-SPV-011infected cell lysate, (G) S-SPV-012 infected cell lysate, (H) S-SPV-013infected cell lysate. Cell lysates are prepared as described in thePREPARATION OF INFECTED CELL LYSATES. Approximately 1/5 of the totallysates sample is loaded in each lane.

FIG. 17

Map showing the 3,628 base pair BglII to HindIII swinepox virus DNAfragment inserted into homology vector 515-85.1.

Two open reading frames, O2L and O1L, are shown with the number of aminoacids coding in each open reading frame. The homology vector 738-94.5contains a deletion of SPV DNA from nucleotides 1381 to 2133 (SEQ ID NO.113) . The lacZ gene has been inserted into this region and is expressedfrom the O1L promoter. Positions of restriction sites AccI, BglII, andHindIII are shown.

FIGS. 18A, 18B, 18C and 18D show a detailed description of SwinepoxVirus S-SPV-034 and the DNA insertion in Homology Vector 723-59A9.22.FIG. 18A contains a diagram showing the orientation of DNA fragmentsassembled in plasmid 723-59A9.22 and a table indicating the origin ofeach fragment. FIG. 18B showns the sequences located at Junctions A andB between fragments, FIG. 18C shows the sequences located at Junction C,and FIG. 18D shows the sequences located at Junctions D and E. Therestriction sites used to generate each fragment as well as syntheticlinker sequences which are used to join the fragments are described foreach junction in FIGS. 18B to 18D. The location of several gene codingregions and regulatory elements is also given. The following twoconventions are used: numbers in parentheses, ( ), refer to amino acids,and restriction sites in brackets, [ ], indicate the remnants of siteswhich are destroyed during construction. The following abbreviations areused: swinepox virus (SPV), equine influenza virus (EIV), Escherichiacoli (E. coli), pox synthetic late promoter 1 (LP1), pox synthetic latepromoter 2 early promoter 2 (LP2EP2), neuraminidase (NA), Prague (PR),polymerase chain reaction (PCR), base pairs (BP).

FIGS. 19A, 19B, 19C and 19D show a detailed description of SwinepoxVirus S-SPV-015 and the DNA insertion in Homology Vector 727-54.60. FIG.19A contains a diagram showing the orientation of DNA fragmentsassembled in plasmid 727-54.60 and a table indicating the origin of eachfragment. FIG. 19B shows the sequences located at Junctions A and Bbetween fragments, FIG. 19C shows the sequences located at Junction C,and FIG. 19D shows the sequences located at Junctions D and E. Therestriction sites used to generate each fragment as well as syntheticlinker sequences which are used to join the fragments are described foreach junction in FIGS. 19B to 19D. The location of several gene codingregions and regulatory elements is also given. The following twoconventions are used: numbers in parentheses, ( ), refer to amino acids,and restriction sites in brackets, [ ], indicate the remnants of siteswhich are destroyed during construction. The following abbreviations areused: swinepox virus (SPV), pseudorabies virus (PRV), Escherichia coli(E. coli), pox synthetic late promoter 1 (LP1), pox synthetic latepromoter 2 early promoter 2 (LP2EP2), glycoprotein B (gB), base pairs(BP).

FIGS. 20A, 20B, 20C, and 20D show a detailed description of SwinepoxVirus S-SPV-031 and the DNA insertion in Homology Vector 727-67.18. FIG.20A contains a diagram showing the orientation of DNA fragmentsassembled in plasmid 727-67.18 and a table indicating the origin of eachfragment. FIG. 20B shows the sequences located at Junctions A and Bbetween fragments, FIG. 20C shows the sequences located at Junction C,and FIG. 20D shows the sequences located at Junctions D and E. Therestriction sites used to generate each fragment as well as syntheticlinker sequences which are used to join the fragments are described foreach junction in FIGS. 20B to 20D. The location of several gene codingregions and regulatory elements is also given.

The following two conventions are used: numbers in parentheses, ( ),refer to amino acids, and restriction sites in brackets, [ ], indicatethe remnants of sites which are destroyed during construction. Thefollowing abbreviations are used: swinepox virus (SPV), Escherichia coli(E. coli), pox synthetic late promoter 1 (LP1), pox synthetic earlypromoter 1 late promoter 2 (EP1LP2), antigen (Ag), base pairs (BP).

FIGS. 21A, 21B, 21C and 21D show a detailed description of SwinepoxVirus S-SPV-033 and the DNA insertion in Homology Vector 732-18.4. FIG.21A contains a diagram showing the orientation of DNA fragmentsassembled in plasmid 732-18.4 and a table indicating the origin of eachfragment. FIG. 21B shows the sequences located at Junctions A and Bbetween fragments, FIG. 21C shows the sequences located at Junction C,and FIG. 21D shows the sequences located at Junctions D and E. Therestriction sites used to generate each fragment as well as syntheticlinker sequences which are used to join the fragments are described foreach junction in FIGS. 21B to 21D. The location of several gene codingregions and regulatory elements is also given. The following twoconventions are used: numbers in parentheses, ( ), refer to amino acids,and restriction sites in brackets, [ ], indicate the remnants of siteswhich are destroyed during construction. The following abbreviations areused: swinepox virus (SPV), Escherichia coli (E. coli), pox syntheticlate promoter 1 (LP1), pox synthetic late promoter 2 early promoter 2(LP2EP2), equine influenza virus (EIV), neuraminidase (NA), Alaska (AK),polymerase chain reaction (PCR), base pairs (BP).

FIGS. 22A, 22B and 22C show a detailed description of Swinepox VirusS-SPV-036 and the DNA insertion in Homology Vector 741-80.3. FIG. 22Acontains a diagram showing the orientation of DNA fragments assembled inplasmid 741-80.3 and a is table indicating the origin of each fragment.FIG. 22B shows the sequences located at Junctions A, B, and C betweenfragments and FIG. 22C shows the sequences located at Junctions D, E andF. The restriction sites used to generate each fragment as well assynthetic linker sequences which are used to join the fragments aredescribed for each junction in FIGS. 22B and 22C. The location ofseveral gene coding regions and regulatory elements is also given. Thefollowing two conventions are used: numbers in parentheses, ( ), referto amino acids, and restriction sites in brackets, [ ], indicate theremnants of sites which are destroyed during construction. The followingabbreviations are used: swinepox virus (SPV), pseudorabies virus (PRV),Escherichia coli (E. coli), human cytomegalovirus immediate early (HCMVIE), pox synthetic late promoter 1 (LP1), pox synthetic late promoter 2early promoter 2 (LP2EP2), polyadenylation site (poly A), base pairs(BP).

FIGS. 23A, 23B, 23C and 23D show a detailed description of SwinepoxVirus S-SPV-035 and the DNA insertion in Homology Vector 741-84.14. FIG.23A contains a diagram showing the orientation of DNA fragmentsassembled in plasmid 741-84.14 and a table indicating the origin of eachfragment. FIG. 23B shows the sequences located at Junctions A and Bbetween fragments, FIG. 23C shows the sequences located at Junction C,and FIG. 23D shows the sequences located at Junctions D and E. Therestriction sites used to generate each fragment as well as syntheticlinker sequences which are used to join the fragments are described foreach junction in FIGS. 23B to 23D. The location of several gene codingregions and regulatory elements is also given. The following twoconventions are used: numbers in parentheses, ( ), refer to amino acids,and restriction sites in brackets, [ ], indicate the remnants of siteswhich are destroyed during construction. The following abbreviations areused: swinepox virus (SPV), pseudorabies virus (PRV), Escherichia coli(E. coli), pox synthetic late promoter 1 (LP1), pox synthetic latepromoter 2 early promoter 2 (LP2EP2), interleukin-2 (IL-2), glycoproteinX (gX) polymerase chain reaction (PCR), sequence (seq), base pairs (BP).

FIGS. 24A, 24B, 24C and 24D show a detailed description of SwinepoxVirus S-SPV-038 and the DNA insertion in Homology Vector 744-34. FIG.24A contains a diagram showing the orientation of DNA fragmentsassembled in plasmid 744-34 and a table indicating the origin of eachfragment. FIG. 24B shows the sequences located at Junction A and Bbetween fragments, FIG. 24C shows the sequences located at Junction C,and FIG. 24D shows the sequences located at Junctions D and E. Therestriction sites used to generate each fragment as well as syntheticlinker sequences which are used to join the fragments are described foreach junction in FIGS. 24B and 24D. The location of several gene codingregions and regulatory elements is also given.

The following two conventions are used: numbers in parentheses, ( ),refer to amino acids, and restriction sites in brackets, [ ], indicatethe remnants of sites which are destroyed during construction. Thefollowing abbreviations are used: swinepox virus (SPV), equineherpesvirus type 1 (EHV-1) , Escherichia coli (E. coli) , pox syntheticlate promoter 1 (LPs), pox synthetic late promoter 2 early promoter 2(LP2EP2), glycoprotein B (gB), polymerase chain reaction (PCR), basepairs (BP).

FIGS. 25A, 25B, 25C, and 25D show a detailed description of SwinepoxVirus S-SPV-039 and the DNA insertion in Homology Vector 744-38. FIG.25A contains a diagram showing the orientation of DNA fragmentsassembled in plasmid 744-38 and a table indicating the origin of eachfragment. FIG. 25B shows the sequences located at Junction A and Bbetween fragments. FIG. 25C shows the sequences located at Junction Cand FIG. 25D shows the sequences located at Junctions D and E. Therestriction sites used to generate each fragment as well as syntheticlinker sequences which are used to join the fragments are described foreach junction in FIGS. 25B to 25D. The location of several gene codingregions and regulatory elements is also given. The following twoconventions are used: numbers in parentheses, ( ), refer to amino acids,and restriction sites in brackets, [ ], indicate the remnants of siteswhich are destroyed during construction. The following abbreviations areused: swinepox virus (SPV), equine herpesvirus type 1 (EHV-1),Escherichia coli (E. coli), pox synthetic late promoter 1 (LP1), poxsynthetic late promoter 2 early promoter 2 (LP2EP2), glycoprotein D(gD), polymerase chain reaction (PCR), base pairs (BP).

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a recombinant swinepox virus (SPV)capable of replication in an animal into which the recombinant swinepoxvirus is introduced which comprises swinepox viral DNA and foreign DNAencoding RNA which does not naturally occur in the animal into which therecombinant swinepox virus is introduced, the foreign DNA being insertedinto the swinepox viral DNA at an insertion site which is not essentialfor replication of the swinepox virus and being under the control of apromoter.

For purposes of this invention , "a recombinant swinepox virus capableof replication" is a live swinepox virus which has been generated by therecombinant methods well known to those of skill in the art, e.g., themethods set forth in HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATINGRECOMBINANT SPV in Materials and Methods and has not had geneticmaterial essential for the replication of the recombinant swinepox virusdeleted.

For purposes of this invention, "an insertion site which is notessential for replication of the swinepox virus" is a location in theswinepox viral genome where a sequence of DNA is not necessary for viralreplication, for example, complex protein binding sequences, sequenceswhich code for reverse transcriptase or an essential glycoprotein, DNAsequences necessary for packaging, etc.

For purposes of this invention, a "promoter" is a specific DNA sequenceon the DNA molecule to which the foreign RNA polymerase attaches and atwhich transcription of the foreign RNA is initiated.

The invention further provides foreign RNA which encodes a polypeptide.Preferably, the polypeptide is antigenic in the animal. Preferably, thisantigenic polypeptide is a linear polymer of more than 10 amino acidslinked by peptide bonds which stimulates the animal to produceantibodies.

The invention further provides an insertion site present within thelarger HindIII to BglII subfragment of the HindIII M fragment ofswinepox viral DNA. Preferably, the insertion site is within an openreading frame contained in the HindIII to BctlII subfragment.Preferably, the insertion site is the AccI restriction endonuclease sitelocated in the HindIII to BclII subfragment.

The invention further provides an insertion site within an open readingframe encoding swinepox thymidine kinase. Preferably, the insertion siteis the NdeI restriction endonuclease site located within the swinepoxvirus thymidine kinase gene.

For purposes of this invention, an "open reading frame" is a segment ofDNA which contains codons that can be transcribed into RNA which can betranslated into an amino acid sequence and which does not contain atermination codon.

The invention further provides a recombinant swinepox virus capable ofreplication which contains a foreign DNA encoding a polypeptide which isa detectable marker. Preferably the detectable marker is the polypeptideE. coli β-galactosidase. Preferably, the insertion site for the foreignDNA encoding E. coli β-galactosidase is the AccI restrictionendonuclease site located within the HindIII M fragment of the swinepoxviral DNA. Preferably, this recombinant swinepox virus is designatedS-SPV-003 (ATCC Accession No. VR 2335). The S-SPV-003 swinepox virus hasbeen deposited pursuant to the Budapest Treaty on the InternationalDeposit of Microorganisms for the Purposes of Patent Procedure with thePatent Culture Depository of the American Type Culture Collection, 12301Parklawn Drive, Rockville, Md. 20852 U.S.A. under ATCC Accession No. VR2335.

For purposes of this invention, a "polypeptide which is a detectablemarker" includes the bimer, trimer and tetramer form of the polypeptide.E. coli β-galactosidase is a tetramer composed of four polypeptides ormonomer sub-units.

The invention further provides a recombinant swinepox virus capable ofreplication which contains foreign DNA encoding an antigenic polypeptidewhich is or is from pseudorabies virus (PRV) g50 (gpD), pseudorabiesvirus (PRV) II (gpB), Pseudorabies virus (PRV) gIII (gpC), pseudorabiesvirus (PRV) glycoprotein H, pseudorabies virus (PRV) glycoprotein E,Transmissible gastroenteritis (TGE) glycoprotein 195, Transmissiblegastroenteritis (TGE) matrix protein, swine rotavirus glycoprotein 38,swine parvovirus capsid protein, Serpulina hydodysenteriae protectiveantigen, Bovine Viral Diarrhea (BVD) glycoprotein 55, Newcastle DiseaseVirus (NDV) hemagglutinin-neuraminidase, swine flu hemagglutinin orswine flu neuraminidase. Preferably, the antigenic polypeptide isPseudorabies Virus (PRV) g50 (gpD). Preferably, the antigenic protein isNewcastle Disease Virus (NDV) hemagglutinin-neuraminidase.

The invention further provides a recombinant swinepox virus capable ofreplication which contains foreign DNA encoding an antigenic polypeptidewhich is or is from Serpulina hyodysenteriae, Foot and Mouth DiseaseVirus, Hog Cholera Virus, Swine Influenza Virus, African Swine FeverVirus or Mycoplasma hyopneumoniae.

The invention further provides for a recombinant swinepox virus capableof replication which contains foreign DNA encoding pseudorabies virus(PRV) g50 (gpD). This recombinant swinepox virus can be furtherengineered to contain foreign DNA encoding a detectable marker, such asE. coli B-galactosidase. A preferred site within the swinepox viralgenome for insertion of the foreign DNA encoding PRV g50 (gpD) and E.coli B-galactosidase is the AccI site within the HindIII M fragment ofthe swinepox viral DNA. Preferably, this recombinant swinepox virus isdesignated S-SPV-008 (ATCC Accession No. VR 2339). The S-SPV-008swinepox virus has been deposited pursuant to the Budapest Treaty on theInternational Deposit of Microorganisms for the Purposes of PatentProcedure with the Patent Culture is Depository of the American TypeCulture Collection, 12301 Parklawn Drive, Rockville, Md. 20852 U.S.A.under ATCC Accession No. VR 2339.

The invention further provides for a recombinant swinepox virus capableof replication which contains foreign DNA encoding pseudorabies virus(PRV) gIII (gpC). This recombinant swinepox virus can also be furtherengineered to contain foreign DNA encoding a detectable marker, such asE. coli B-galactosidase. A preferred site within the swinepox viral DNAfor insertion of the foreign DNA encoding PRV C gene and E. coliB-galactosidase is the AccI site within the HindIII M fragment of theswinepox viral DNA. Preferably, this recombinant swinepox virus isdesignated S-SPV-011, S-SPV-012, or S-SPV-013. The swinepox virusdesignated S-SPV-013 has been deposited pursuant to the Budapest Treatyon the International Deposit of Microorganisms for the Purposes ofPatent Procedure with the Patent Culture Depository of the American TypeCulture Collection, 12301 Parklawn Drive, Rockville, Md. 20852 U.S.A.

The invention further provides for a recombinant swinepox virus capableof replication which contains foreign DNA encoding pseudorabies virus(PRV) gII (gpB). This recombinant swinepox virus can also be furtherengineered to contain foreign DNA encoding a detectable marker, such asE. coli B-galactosidase. A preferred site within the swinepox viral DNAfor insertion of the foreign DNA encoding PRV gII (gpB) and E. coliB-galactosidase is the AccI site within the HindIII M fragment of theswinepox viral DNA. Preferably, this recombinant swinepox virus isdesignated S-SPV-015. The S-SPV-015 swinepox virus has been deposited onJul. 22, 1994 pursuant to the Budapest Treaty on the InternationalDeposit of Microorganisms for the Purposes of Patent Procedure with thePatent Culture Depository of the American Type Culture Collection, 12301Parklawn Drive, Rockville, Md. 20852 U.S.A.

The invention further provides for a recombinant swinepox virus capableof replication which contains foreign DNA encoding pseudorabies virus(PRV) g50 (gpD) and foreign DNA encoding pseudorabies virus (PRV) gIII(gpC). This recombinant swinepox virus can also be further engineered tocontain foreign DNA encoding a detectable marker, such as E. coliB-galactosidase. A preferred site within the swinepox viral DNA forinsertion of the foreign DNA encoding PRV g50 (gpD), PRV gIII (gpC) andE. coli B-galactosidase is the AccI site within the HindIII M fragmentof the swinepox viral DNA.

The invention further provides for a recombinant swinepox virus capableof replication which contains foreign DNA encoding pseudorabies virus(PRV) g50 (gpD) and foreign DNA encoding pseudorabies virus (PRV) gII(gpB). This recombinant swinepox virus can also be further engineered tocontain Foreign DNA encoding a detectable marker, such as E. coliB-galactosidase. A preferred site within the swinepox viral genome forinsertion of foreign DNA encoding PRV g50 (gpD), PRV gII (gpB) and E.coli B-galactosidase is the AccI site within the HindIII M fragment ofthe swinepox viral DNA.

The invention further provides for a recombinant swinepox virus capableof replication which contains foreign DNA encoding pseudorabies virus(PRV) gIII (gpC) and foreign DNA encoding pseudorabies virus (PRV) gII(gpB). This recombinant swinepox virus can also be further engineered tocontain foreign DNA encoding a detectable marker, such as E. coliB-galactosidase. A preferred site within the swinepox viral genome forinsertion of foreign DNA encoding PRV gIII (gpC), PRV gIl (gpB) and E.coli B-galactosidase is the AccI site within the HindIII M fragment ofthe swinepox viral DNA.

The invention further provides for a recombinant swinepox virus capableof replication which contains foreign DNA encoding pseudorabies virus(PRV) g50 (gpD), foreign DNA encoding pseudorabies virus (PRV) gIII(gpC), and foreign DNA encoding pseudorabies virus (PRV) gII (gpB). Thisrecombinant swinepox virus can also be further engineered to containforeign DNA encoding a detectable marker, such as E. coliB-galactosidase.

A preferred site within the swinepox viral genome for insertion offoreign DNA encoding PRV g50 (gpD), PRV gIII (gpC), PRV gII (gpB) and E.coli B-galactosidase is the AccI site within the HindIII M fragment ofthe swinepox viral DNA.

The invention further provides for a recombinant swinepox virus capableof replication which contains foreign DNA encoding RNA encoding theantigenic polypeptide Newcastle Disease Virus (NDV)hemagglutinin-neuraminidase further comprising foreign DNA encoding apolypeptide which is a detectable marker. Preferably, this recombinantswinepox virus is designated S-SPV-009 (ATCC Accession No. VR 2344). TheS-SPV-009 swinepox virus has been deposited pursuant to the BudapestTreaty on the International Deposit of Microorganisms for the Purposesof Patent Procedure with the Patent Culture Depository of the AmericanType Culture Collection, 12301 Parklawn Drive, Rockville, Md. 20852U.S.A. under ATCC Accession No. VR 2344.

The present invention further provides a recombinant swinepox viruswhich comprises a foreign DNA sequence inserted into a non-essentialsite of the swinepox genome, wherein the foreign DNA sequence encodes anantigenic polypeptide derived from infectious bovine rhinotracheitisvirus and is capable of being expressed in a host infected by therecombinant swinepox virus. Examples of such antigenic polypeptide areinfectious bovine rhinotracheitis virus glycoprotein E and glycoproteinG. Preferred embodiment of this invention are recombinant swinepoxviruses designated S-SPV-017 and S-SPV-019.

The present invention further provides a recombinant swinepox viruswhich comprises a foreign DNA sequence inserted into a non-essentialsite of the swinepox genome, wherein the foreign DNA sequence encodes anantigenic polypeptide derived from infectious laryngotracheitis virusand is capable of being expressed in a host infected by the recombinantswinepox virus. Examples of such antigenic polypeptide are infectiouslaryngotracheitis virus glycoprotein G and glycoprotein I. Preferredembodiment of this invention are recombinant swinepox viruses designatedS-SPV-014 and S-SPV-016.

The present invention further provides a recombinant swinepox viruswhich comprises a foreign DNA sequence inserted into a non-essentialsite of the swinepox genome, wherein the foreign DNA sequence encodes anantigenic polypeptide derived from a human pathogen and is capable ofbeing expressed in a host infected by the recombinant swinepox virus.

For example, the antigenic polypeptide of a human pathogen is derivedfrom human herpesvirus, herpes simplex virus-1, herpes simplex virus-2,human cytomegalovirus, Epstein-Barr virus, Varicell-Zoster virus, humanherpesvirus-6, human herpesvirus-7, human influenza, humanimmunodeficiency virus, rabies virus, measles virus, hepatitis B virusand hepatitis C virus. Furthermore, the antigenic polypeptide of a humanpathogen may be associated with malaria or malignant tumor from thegroup conisting of Plasmodium falciparum, Bordetella pertusis, andmalignant tumor.

In one embodiment of the invention, a recombinant swinepox viruscontains the foreign DNA sequence encoding hepatitis B virus coreprotein. Preferably, such virus recombinant virus is designatedS-SPV-031.

The present invention further provides a recombinant swinepox viruswhich comprises a foreign DNA sequence inserted into a non-essentialsite of the swinepox genome, wherein the foreign DNA sequence encodes acytokine capable of stimulating an immune in a host infected by therecombinant swinepox virus and is capable of being expressed in the hostinfected.

For example, the cytokine can be, but not limited to, interleukin-2,interleukin-6, interleukin-12, interferons, granulocyte-macrophagecolony stimulating factors, and interleukin receptors.

In one embodiment of the invention, a recombinant swinepox viruscontains a foreign DNA sequence encoding human interleukin-2.Preferably, such recombinant virus is designated S-SPV-035.

The present invention further provides a recombinant swinepox viruswhich comprises a foreign DNA sequence inserted into a non-essentialsite of the swinepox genome, wherein the foreign DNA sequence encodes anantigenic polypeptide derived from an equine pathogen and is capable ofbeing expressed in a host infected by the recombinant swinepox virus.

The antigenic polypeptide of an equine pathogen can derived from equineinfluenza virus or equine herpesvirus. Examples of such antigenicpolypeptide are equine influenza virus type A/Alaska 91 neuraminidase,equine influenza virus type A/Prague 56 neuraminidase, equine influenzavirus type A/Miami 63 neuraminidase, equine influenza virus typeA/Kentucky 81 neuraminidaseequine herpesvirus type 1 glycoprotein B, andequine herpesvirus type 1 glycoprotein D. Preferred embodiments of suchrecombinant virus are designated S-SPV-033, S-SPV-034, S-SPV-038, andS-SPV-039.

The present invention further provides a recombinant swinepox viruswhich comprises a foreign DNA sequence inserted into a non-essentialsite of the swinepox genome, wherein the foreign DNA sequence encodes anantigenic polypeptide derived from bovine respiratory syncytial virus orbovine parainfluenza virus, and is capable of being expressed in a hostinfected by the recombinant swinepox virus.

For example, the antigenic polypeptide of derived from bovinerespiratory syncytial virus equine pathogen can derived from equineinfluenza virus is bovine respiratory syncytial virus attachment protein(BRSV G), bovine respiratory syncytial virus fusion protein (BRSV F),bovine respiratory syncytial virus nucleocapsid protein (BRSV N), bovineparainfluenza virus type 3 fusion protein, and the bovine parainfluenzavirus type 3 hemagglutinin neuraminidase.

Preferred embodiments of a recombinant virus containing a foreign DNAencoding an antigenic polypeptide from a bovine respiratory syncytialvirus are designated S-SPV-020, S-SPV-029, and S-SPV-030.

And a preferred embodiment of a recombinant virus containing a foreignDNA encoding an antigenic polypeptide from a bovine parainfluenza virusare designated S-SPV-028.

The present invention further provides a recombinant swinepox viruswhich comprises a foreign DNA sequence inserted into a non-essentialsite of the swinepox genome, wherein the foreign DNA sequence encodesbovine viral diarrhea virus glycoprotein 48 or glycoprotein 53, andwherein the foreign DNA sequence is capable of being expressed in a hostinfected by the recombinant swinepox virus. Preferred embodiments ofsuch virus are designated S-SPV-032 and S-SPV-040.

The present invention further provides a recombinant swinepox viruswhich comprises a foreign DNA sequence inserted into a non-essentialsite of the swinepox genome, wherein the foreign DNA sequence encodes anantigenic polypeptide derived from infectious bursal disease virus andwherein the foreign DNA sequence is capable of being expressed in a hostinfected by the recombinant swinepox virus. Examples of such antigenicpolypeptide are infectious bursal disease virus polyprotein and VP2.Preferred embodiments of such virus are designated S-SPV-026 andS-SPV-027.

The invention further provides that the inserted foreign DNA sequence isunder the control of a promoter. Preferably, the promoter is a swinepoxviral promoter. Preferably, the promoter is a synthetic pox viralpromoter. For purposes of this invention, the promoters were generatedby methods well known to those of skill in the art, for example, as setforth in the STRATEGY FOR THE CONSTRUCTION OF SYNTHETIC POX VIRALPROMOTERS in Materials and Methods. For purposes of this invention, asynthetic pox promoter includes a synthetic late pox promoter, asynthetic early pox promoter or a synthetic early/late pox promoter.

The invention provides for a homology vector for producing a recombinantswinepox virus by inserting foreign DNA into the genomic DNA of aswinepox virus. The homology vector comprises a double-stranded DNAmolecule consisting essentially of a double-stranded foreign DNAencoding RNA which does not naturally occur in an animal into which therecombinant swinepox virus is introduced, with at one end of the foreignDNA, double-stranded swinepox viral DNA homologous to genomic DNAlocated at one side of a site on the genomic DNA which is not essentialfor replication of the swinepox virus, and at the other end of theforeign DNA, double-stranded swinepox viral DNA homologous to genomicDNA located at the other side of the same site on the genomic DNA.Preferably, the RNA encodes a polypeptide.

In one embodiment, the polypeptide is a detectable marker. Preferably,the polypeptide which is a detectable marker is E. coli β-galactosidase.

In one embodiment, the polypeptide is antigenic in the animal.Preferably, the antigenic polypeptide is or is from pseudorabies virus(PRV) g50 (gpD), pseudorabies virus (PRV) gII (gpB), Pseudorabies virus(PRV) gIII (gpC), Pseudorabies virus (PRV) glycoprotein H, Transmissiblegastroenteritis (TGE) glycoprotein 195, Transmissible gastroenteritis(TGE) matrix protein, swine rotavirus glycoprotein 38, swine parvoviruscapsid protein, Serpulina hydodysenteriae protective antigen, BovineViral Diarrhea (BVD) glycoprotein 55, Newcastle Disease Virus (NDV)hemagglutinin-neuraminidase, swine flu hemagglutinin or swine fluneuraminidase. Preferably, the antigenic polypeptide is or is fromSerpulina hyodesenteriae, Foot and Mouth Disease Virus, Hog CholeraVirus, Swine Influenza Virus, African Swine Fever Virus or Mycoplasmahyopneumoniae.

In an embodiment of the present invention, the double stranded foreignDNA sequence in the homology vector encodes an antigenic polypeptidederived from a human pathogen.

For example, the antigenic polypeptide of a human pathogen is derivedfrom human herpesvirus, herpes simplex virus-1, herpes simplex virus-2,human cytomegalovirus, Epstein-Barr virus, Varicell-Zoster virus, humanherpesvirus-6, human herpesvirus-7, human influenza, humanimmunodeficiency virus, rabies virus, measles virus, hepatitis B virusand hepatitis C virus. Furthermore, the antigenic polypeptide of a humanpathogen may be associated with malaria or malignant tumor from thegroup conisting of Plasmodium falciparum, Bordetella pertusis, andmalignant tumor.

In an embodiment of the present invention, the double stranded foreignDNA sequence in the homology vector encodes a cytokine capable ofstimulating human immune response. For example, the cytokine can be, butnot limited to, interleukin-2, interleukin-6, interleukin-12,interferons, granulocyte-macrophage colony stimulating factors, andinterleukin receptors.

In an embodiment of the present invention, the double stranded foreignDNA sequence in the homology vector encodes an antigenic polypeptidederived from an equine pathogen.

The antigenic polypeptide of an equine pathogen can derived from equineinfluenza virus or equine herpesvirus. Examples of such antigenicpolypeptide are equine influenza virus type A/Alaska 91 neuraminidase,equine influenza virus type A/Prague 56 neuraminidase, equine influenzavirus type A/Miami 63 neuraminidase, equine influenza virus typeA/Kentucky 81 neuraminidaseequine herpesvirus type 1 glycoprotein B, andequine herpesvirus type 1 glycoprotein D.

In an embodiment of the present invention, the double stranded foreignDNA sequence of the homology vector encodes an antigenic polypeptidederived from bovine respiratory syncytial virus or bovine parainfluenzavirus.

For example, the antigenic polypeptide of derived from bovinerespiratory syncytial virus equine pathogen can derived from equineinfluenza virus is bovine respiratory syncytial virus attachment protein(BRSV G), bovine respiratory syncytial virus fusion protein (BRSV F),bovine respiratory syncytial virus nucleocapsid protein (BRSV N), bovineparainfluenza virus type 3 fusion protein, and the bovine parainfluenzavirus type 3 hemagglutinin neuraminidase.

In an embodiment of the present invention, the double stranded foreignDNA sequence of the homology vector encodes an antigenic polypeptidederived from infectious bursal disease virus. Examples of such antigenicpolypeptide are infectious bursal disease virus polyprotein andinfectious bursal disease virus VP2.

In another embodiment of the present invention, the double-strandedswinepox viral DNA of the homology vectors described above is homologousto genomic DNA present within the larger HindIII to BglII subfragment ofthe HindIII M fragment of swinepox virus. Preferably, thedouble-stranded swinepox viral DNA is homologous to genomic DNA presentwithin the open reading frame contained in this HindIII to BclIIsubfragment. Preferably, the double-stranded swinepox viral DNA ishomologous to genomic DNA present within the AccI restrictionendonuclease site located in this HindIII to BclII subfragment.

For purposes of this invention, a "homology vector" is a plasmidconstructed to insert foreign DNA in a specific site on the genome of aswinepox virus.

In one embodiment of the invention, the double-stranded swinepox viralDNA of the homology vectors described above is homologous to genomic DNApresent within the open reading frame encoding swinepox thymidinekinase. Preferably, the double-stranded swinepox viral DNA is homologousto genomic DNA present within the NdeI restriction endonuclease sitelocated in the open reading frame encoding swinepox thymidine kinase.

The invention further provides a homology vectors described above, theforeign DNA sequence of which is under control of a promoter locatedupstream of the foreign DNA sequence. The promoter can be an endogenousswinepox viral promoter or an exogenous promoter. The promoter can be asynthetic pox viral promoter or human cytomegalovrus immediate earlygene promoter.

The invention further provides a vaccine which comprises an effectiveimmunizing amount of a recombinant swinepox virus of the presentinvention and a suitable carrier.

Suitable carriers for the pseudorabies virus are well known in the artand include proteins, sugars, etc. One example of such a suitablecarrier is a physiologically balanced culture medium containing one ormore stabilizing agents such as stabilized, hydrolyzed proteins,lactose, etc.

For purposes of this invention, an "effective immunizing amount" of therecombinant swinepox virus of the present invention is within the rangeof 10³ to 10⁹ PFU/dose.

The present invention also provides a method of immunizing an animal,wherein the animal is a human, swine, bovine, equine, caprine or ovine.For purposes of this invention, this includes immunizing the animalagainst the virus or viruses which cause the disease or diseasespseudorabies, transmissible gastroenteritis, swine rotavirus, swineparvovirus, Serpulina hyodysenteriae, bovine viral diarrhea, Newcastledisease, swine flu, foot and mouth disease, hog cholera, African swinefever or Mycoplasma hyopneumoniae. For purposes of this invention, themethod of immunizing also includes immunizing the animal against humanpathogens, bovine pathogens, equine pathogens, avian pathogens describedin the preceding part of this section.

The method comprises administering to the animal an effective immunizingdose of the vaccine of the present invention. The vaccine may beadministered by any of the methods well known to those skilled in theart, for example, by intramuscular, subcutaneous, intraperitoneal orintravenous injection. Alternatively, the vaccine may be administeredintranasally or orally.

The present invention also provides a method for testing a swine todetermine whether the swine has been vaccinated with the vaccine of thepresent invention, particularly the embodiment which contains therecombinant swinepox virus S-SPV-008 (ATCC Accession No. VR 2339), or isinfected with a naturally-occurring, wild-type pseudorabies virus. Thismethod comprises obtaining from the swine to be tested a sample of asuitable body fluid, detecting in the sample the presence of antibodiesto pseudorabies virus, the absence of such antibodies indicating thatthe swine has been neither vaccinated nor infected, and for the swine inwhich antibodies to pseudorabies virus are present, detecting in thesample the absence of antibodies to pseudorabies virus antigens whichare normally present in the body fluid of a swine infected by thenaturally-occurring pseudorabies virus but which are not present in avaccinated swine indicating that the swine was vaccinated and is notinfected.

The present invention also provides a host cell infected with arecombinant swinepox virus capable of replication. In one embodiment,the host cell is a mammalian cell. Preferably, the mammalian cell is aVero cell. Preferably, the mammalian cell is an ESK-4 cell, PK-15 cellor EMSK cell.

For purposes of this invention a "host cell" is a cell used to propagatea vector and its insert. Infecting the cells was accomplished by methodswell known to those of skill in the art, for example, as set forth inINFECTION--TRANSFECTION PROCEDURE in Material and Methods.

Methods for constructing, selecting and purifying recombinant swinepoxviruses described above are detailed below in Materials and Methods.

Materials and Methods

PREPARATION OF SWINEPOX VIRUS STOCK SAMPLES. Swinepox virus (SPV)samples were prepared by infecting embryonic swine kidney (EMSK) cells,ESK-4 cells, PK-15 cells or Vero cells at a multiplicity of infection of0.01 PFU/cell in a 1:1 mixture of Iscove's Modified Dulbecco's Medium(IMDM) and RPMI 1640 medium containing 2 mM glutamine, 100 units/mlpenicillin, 100 units/ml streptomycin (these components were obtainedfrom Sigma or equivalent supplier, and hereafter are referred to as EMSKnegative medium). Prior to infection, the cell monolayers were washedonce with EMSK negative medium to remove traces of fetal bovine serum.The SPV contained in the initial inoculum (0.5 ml for 10 cm plate; 10 mlfor T175 cm flask) was then allowed to absorb onto the cell monolayerfor two hours, being redistributed every half hour. After this period,the original inoculum was brought up to the recommended volume with theaddition of complete EMSK medium (EMSK negative medium plus 5% fetalbovine serum). The plates were incubated at 37° C. in 5% CO₂ untilcytopathic effect was complete. The medium and cells were harvested andfrozen in a 50 ml conical screw cap tube at -70° C. Upon thawing at 37°C., the virus stock was aliquoted into 1.0 ml vials and refrozen at -70°C. The titers were usually about 10⁶ PFU/ml.

PREPARATION OF SPV DNA. For swinepox virus DNA isolation, a confluentmonolayer of EMSK cells in a T175 cm² flask was infected at amultiplicity of 0.1 and incubated 4-6 days until the cells were showing100% cytopathic effect. The infected cells were then harvested byscraping the cells into the medium and centrifuging at 3000 rpm for 5minutes in a clinical centrifuge. The medium was decanted, and the cellpellet was gently resuspended in 1.0 ml Phosphate Buffer Saline (PBS:1.5 g Na₂ HPO₄, 0.2 g KH₂ PO₄, 0.8 g NaCL and 0.2 g KCl per liter H₂ O)(per T175) and subjected to two successive freeze-thaws (-70° C. to 37°C.). Upon the last thaw, the cells (on ice) were sonicated two times for30 seconds each with 45 seconds cooling time in between. Cellular debriswas then removed by centrifuging (Sorvall RC-5B superspeed centrifuge)at 3000 rpm for 5 minutes in a HB4 rotor at 40° C. SPV virions, presentin the supernatant, were then pelleted by centrifugation at 15,000 rpmfor 20 minutes at 40° C. in a SS34 rotor (Sorvall) and resuspended in mMTris (pH 7.5). This fraction was then layered onto a 36% sucrosegradient (w/v in 10 mM tris pH 7.5) and centrifuged (Beckman L8-70MUltracentrifuge) at 18,000 rpm for 60 minutes in a SW41 rotor (Beckman)at 4° C. The virion pellet was resuspended in 1.0 ml of 10 mM tris pH7.5 and sonicated on ice for 30 seconds. This fraction was layered ontoa 20% to 50% continuous sucrose gradient and centrifuged 16,000 rpm for60 minutes in a SW41 rotor at 4° C. The SPV virion band located aboutthree quarters down the gradient was harvested, diluted with 20% sucroseand pelleted by centrifugation at 18,000 rpm for 60 minutes in a SW41rotor at 4° C. The resultant pellet was then washed once with 10 mM TrispH 7.5 to remove traces of sucrose and finally resuspended in 10 mM TrispH 7.5. SPV DNA was then extracted from the purified virions by lysis (4hours at 60° C.) induced by the addition of EDTA, SDS, and proteinase Kto final concentrations of 20 mM, 0.5% and 0.5 mg/ml, respectively.After digestion, three phenol: chloroform (1:1) extractions wereconducted and the sample precipitated by the addition of two volumes ofabsolute ethanol and incubation at -20° C. for 30 minutes. The samplewas then centrifuged in an Eppendorf minifuge for 5 minutes at fullspeed. The supernatant was decanted, and the pellet air dried andrehydrated in 0.01 M Tris pH 7.5, 1 mM EDTA at 4° C.

PREPARATION OF INFECTED CELL LYSATES. For cell lysate preparation, serumfree medium was used. A confluent monolayer of cells (EMSK, ESK-4, PK-15or Vero for SPV or VERO for PRV) in a 25 cm² flask or a 60 mm petri dishwas infected with 100 μl of virus sample. After cytopathic effect wascomplete, the medium and cells were harvested and the cells werepelleted at 3000 rpm for 5 minutes in a clinical centrifuge. The cellpellet was resuspended in 250 μl of disruption buffer (2% sodium dodecylsulfate, 2% β-mercapto-ethanol). The samples were sonicated for 30seconds on ice and stored at -20° C.

WESTERN BLOTTING PROCEDURE. Samples of lysates and protein standardswere run on a polyacrylamide gel according to the procedure of Laemnli(1970). After gel electrophoresis the proteins were transferred andprocessed according to Sambrook et al. (1982). The primary antibody wasa swine anti-PRV serum (Shope strain; lot370, PDV8201, NVSL, Ames, IA)diluted 1:100 with 5% non-fat dry milk in Tris-sodium chloride, andsodium Azide (TSA: 6.61 g Tris-HCl, 0.97 g Tris-base, 9.0 g NaCl and 2.0g Sodium Azide per liter H₂ O). The secondary antibody was a goatanti-swine alkaline phosphatase conjugate diluted 1:1000 with TSA.

MOLECULAR BIOLOGICAL TECHNIQUES. Techniques for the manipulation ofbacteria and DNA, including such procedures as digestion withrestriction endonucleases, gel electrophoresis, extraction of DNA fromgels, ligation, phosphorylation with kinase, treatment with phosphatase,growth of bacterial cultures, transformation of bacteria with DNA, andother molecular biological methods are described by Maniatis et al.(1982) and Sambrook et al. (1989). Except as noted, these were used withminor variation.

DNA SEQUENCING. Sequencing was performed using the USE Sequenase Kit and³⁵ S-dATP (NEN). Reactions using both the dGTP mixes and the dITP mixeswere performed to clarify areas of compression. Alternatively,compressed areas were resolved on formamide gels. Templates weredouble-stranded plasmid subclones or single stranded M13 subclones, andprimers were either made to the vector just outside the insert to besequenced, or to previously obtained sequence.

Sequence obtained was assembled and compared using Dnastar software.Manipulation and comparison of sequences obtained was performed withSuperclone™ and Supersee™ programs from Coral Software.

CLONING WITH THE POLYMERASE CHAIN REACTION. The polymerase chainreaction (PCR) was used to introduce restriction sites convenient forthe manipulation of various DNAs. The procedures used are described byInnis, et al. (1990). In general, amplified fragments were less than 500base pairs in size and critical regions of amplified fragments wereconfirmed by DNA sequencing. The primers used in each case are detailedin the descriptions of the construction of homology vectors below.

HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. Thismethod relies upon the homologous recombination between the swinepoxvirus DNA and the plasmid homology vector DNA which occurs in the tissueculture cells containing both swinepox virus DNA and transfected plasmidhomology vector. For homologous recombination to occur, the monolayersof EMSK cells are infected with S-SPV-001 (Kasza SPV strain, 17) at amultiplicity of infection of 0.01 PFU/cell to introduce replicating SPV(i.e. DNA synthesis) into the cells. The plasmid homology vector DNA isthen transfected into these cells according to theINFECTION--TRANSFECTION PROCEDURE. The construction of homology vectorsused in this procedure is described below

INFECTION--TRANSFECTION PROCEDURE. 6 cm plates of EMSK cells (about 80%confluent) were infected with S-SPV-001 at a multiplicity of infectionof 0.01 PFU/cell in EMSK negative medium and incubated at 37° C. in ahumidified 5% CO₂ environment for 5 hours. The transfection procedureused is essentially that recommended for Lipofectin™ Reagent (BRL).Briefly, for each 6 cm plate, 15 μg of plasmid DNA was diluted up to 100μl with H₂ O. Separately, 50 micrograms of Lipofectin Reagent wasdiluted to 100 μl with H₂ O. The 100 μl of diluted Lipofectin Reagentwas then added dropwise to the diluted plasmid DNA contained in apolystyrene 5 ml snap cap tube and mixed gently. The mixture was thenincubated for 15-20 minutes at room temperature. During this time, thevirus inoculum was removed from the 6 cm plates and the cell monolayerswashed once with EMSK negative medium. Three ml of EMSK negative mediumwas then added to the plasmid DNA/lipofectin mixture and the contentspipetted onto the cell monolayer. The cells were incubated overnight(about 16 hours) at 37° C. in a humidified 5% CO₂ environment. The nextday the 3 ml of EMSK negative medium was removed and replaced with 5 mlEMSK complete medium. The cells were incubated at 37° C. in 5% CO₂ for3-7 days until cytopathic effect from the virus was 80-100%. Virus washarvested as described above for the preparation of virus stocks. Thisstock was referred to as a transfection stock and was subsequentlyscreened for recombinant virus by the BLUOGAL SCREEN FOR RECOMBINANTSWINEPOX VIRUS OR CPRG SCREEN FOR RECOMBINANT SWINEPOX VIRUS.

SCREEN FOR RECOMBINANT SPV EXPRESSING B-GALACTOSIDASE (BLUOGAL AND CPRGASSAYS). When the E. coli β-galactosidase (lacZ) marker gene wasincorporated into a recombinant virus the plaques containing therecombinants were visualized by one of two simple methods. In the firstmethod, the chemical Bluogal™ (Bethesda Research Labs) was incorporated(200 μg/ml) into the agarose overlay during the plaque assay, andplaques expressing active β-galactosidase turned blue. The blue plaqueswere then picked onto fresh cells (EMSK) and purified by further blueplaque isolation. In the second method, CPRG (Boehringer Mannheim) wasincorporated (400 μg/ml) into the agarose overlay during the plaqueassay, and plaques expressing active β-galactosidase turned red. The redplaques were then picked onto fresh cells (EMSK) and purified by furtherred plaque isolation. In both cases viruses were typically purified withthree rounds of plaque purification.

SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV USING BLACK PLAQUEASSAYS. To analyze expression of foreign antigens expressed byrecombinant swinepox viruses, monolayers of EMSK cells were infectedwith recombinant SPV, overlayed with nutrient agarose media andincubated for 6-7 days at 37° C. for plaque development to occur. Theagarose overlay was then removed from the dish, the cells fixed with100% methanol for 10 minutes at room temperature and the cells airdried. Fixation of the cells results in cytoplasmic antigen as well assurface antigen detection whereas specific surface antigen expressioncan be detected using non-fixed cells. The primary antibody was thendiluted to the appropriate dilution with PBS and incubated on the cellmonolayer for 2 hours at room temperature. To detect PRV g50 (gpD)expression from S-SPV-008, swine anti-PRV serum (Shope strain; lot370,PDV8201, NVSL, Ames, Iowa) was used (diluted 1:100). To detect NDV HNexpression from S-SPV-009, a rabbit antiserum specific for the HNprotein (rabbit anti-NDV#2) was used (diluted 1:1000). Unbound antibodywas then removed by washing the cells three times with PBS at roomtemperature. The secondary antibody, either a goat anti-swine (PRV g50(gpD); S-SPV-008) or goat anti-rabbit (NDV RN; S-SPV-009), horseradishperoxidase conjugate was diluted 1:250 with PBS and incubated with thecells for 2 hours at room temperature. Unbound secondary antibody wasthen removed by washing the cells three times with PBS at roomtemperature. The cells were then incubated 15-30 minutes at roomtemperature with freshly prepared substrate solution (100 μg/ml4-chloro-1-naphthol, 0.003% H₂ O₂ in PBS). Plaques expressing thecorrect antigen stain black.

PROCEDURE FOR PURIFICATION OF VIRAL GLYCOPROTEINS FOR USE ASDIAGNOSTICS. Viral glycoproteins are purified using antibody affinitycolumns. To produce monoclonal antibodies, 8 to 10 week old BALB/cfemale mice are vaccinated intraperitoneally seven times at two to fourweek intervals with 10⁷ PFU of S-SPV-009, -014, -016, -017, -018, or-019. Three weeks after the last vaccination, mice are injectedintraperitoneally with 40 mg of the corresponding viral glycoprotein.Spleens are removed from the mice three days after the last antigendose.

Splenocytes are fused with mouse NS1/Ag4 plasmacytoma cells by theprocedure modified from Oi and Herzenberg, (41). Splenocytes andplasmacytoma cells are pelleted together by centrifugation at 300 × gfor 10 minutes. One ml of a 50% solution of polyethylene glycol (m.w.1300-1600) is added to the cell pellet with stirring over one minute.Dulbecco's modified Eagles's medium (5 ml) is added to the cells overthree minutes. Cells are pelleted by centrifugation at 300 × g for 10minutes and resuspended in medium with 10% fetal bovine serum andcontaining 100 mM hypoxanthine, 0.4 mM aminopterin and 16 mM thymidine(HAT). Cells (100 ml) are added to the wells of eight to ten 96-welltissue culture plates containing 100 ml of normal spleen feeder layercells and incubated at 37° C. Cells are fed with fresh HAT medium everythree to four days.

Hybridoma culture supernatants are tested by the ELISA ASSAY in 96-wellmicrotiter plates coated with 100 ng of viral glycoprotein. Supernatantsfrom reactive hybridomas are further analyzed by black-plaque assay andby Western Blot. Selected hybridomas are cloned twice by limitingdilution. Ascetic fluid is produced by intraperitoneal injection of5×10⁶ hybridoma cells into pristane-treated BALB/c mice.

Cell lysates from S-SPV-009, -014, -016, -017, -018, or -019 areobtained as described in PREPARATION OF INFECTED CELL LYSATES. Theglycoprotein-containing cell lysates (100 mls) are passed through a 2-mlagarose affinity resin to which 20 mg of glycoprotein monoclonalantibody has been immobilized according to manufacturer's instructions(AFC Medium, New Brunswick Scientific, Edison, N.J.). The column iswashed with 100 ml of 0.1% Nonidet P-40 in phosphate-buffered saline(PBS) to remove nonspecifically bound material. Bound glycoprotein iseluted with 100 mM carbonate buffer, pH 10.6 (40). Pre- and postelutedfractions are monitored for purity by reactivity to the SPV monoclonalantibodies in an ELISA system.

ELISA ASSAY. A standard enzyme-linked immunosorbent assay (ELISA)protocol is used to determine the immune status of cattle followingvaccination and challenge.

A glycoprotein antigen solution (100 ml at ng/ml in PBS) is allowed toabsorb to the wells of microtiter dishes for 18 hours at 4° C. Thecoated wells are rinsed one time with PBS. Wells are blocked by adding250 ml of PBS containing 1% BSA (Sigma) and incubating 1 hour at 37° C.The blocked wells are rinsed one time with PBS containing 0.02% Tween20. 50 ml of test serum (previously diluted 1:2 in PBS containing 1%BSA) are added to the wells and incubated 1 hour at 37° C. The antiserumis removed and the wells are washed 3 times with PBS containing 0.02%Tween 20. 50 ml of a solution containing anti-bovine IgG coupled tohorseradish peroxidase (diluted 1:500 in PBS containing 1% BSA,Kirkegaard and Perry Laboratories, Inc.) is added to visualize the wellscontaining antibody against the specific antigen. The solution isincubated 1 hour at 37° C., then removed and the wells are washed 3times with PBS containing 0.02% Tween 20. 100 ml of substrate solution(ATBS, Kirkegaard and Perry Laboratories, Inc.) are added to each welland color is allowed to develop for 15 minutes. The reaction isterminated by addition of 0.1M oxalic acid. The color is read atabsorbance 410 nm on an automatic plate reader.

STRATEGY FOR THE CONSTRUCTION OF SYNTHETIC POX VIRAL PROMOTERS. Forrecombinant swinepox vectors synthetic pox promoters offer severaladvantages including the ability to control the strength and timing offoreign gene expression. We chose to design three promoter cassettesLP1, EP1 and LP2 based on promoters that have been defined in thevaccinia virus (1, 7 and 8). Each cassette was designed to contain theDNA sequences defined in vaccinia flanked by restriction sites whichcould be used to combine the cassettes in any order or combination.Initiator methionines were also designed into each cassette such thatinframe fusions could be made at either EcoRI or BamHI sites. A set oftranslational stop codons in all three reading frames and an earlytranscriptional termination signal (9) were also engineered downstreamof the inframe fusion site. DNA encoding each cassette was synthesizedaccording to standard techniques and cloned into the appropriatehomology vectors (see FIGS. 4, 5 and 8).

VACCINATION STUDIES IN SWINE USING RECOMBINANT SWINEPOX VIRUS CONTAININGPSEUDORABIES VIRUS GLYCOPROTEIN GENES: Young weaned pigs frompseudorabies-free herd are used to test the efficacy of the recombinantswinepox virus containing one or more of the pseudorabies virusglycoprotein genes (SPV/PRV). The piglets are inoculated intradermallyor orally about 10³ to 10⁷ plaque forming units (PFU) of the recombinantSPV/PRV viruses.

Immunity is determined by measuring PRV serum antibody levels and bychallenging the vaccinated pigs with virulent strain of pseudorabiesvirus. Three to four weeks post-vaccination, both vaccinated andnon-vaccinated groups of pigs are challenged with virulent strain ofpseudorabies virus (VDL4892). Post challenge, the pigs are observeddaily for 14 days for clinical signs of pseudorabies.

Serum samples are obtained at the time of vaccination, challenge, and atweekly intervals for two to three weeks post-vaccination and assayed forserum neutralizing antibody.

HOMOLOGY VECTOR 515-85.1. The plasmid 515-85.1 was constructed for thepurpose of inserting foreign DNA into SPV. It contains a unique AccIrestriction enzyme site into which foreign DNA may be inserted. When aplasmid, containing a foreign DNA insert at the AccI site, is usedaccording to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATINGRECOMBINANT SPV a virus containing the foreign DNA will result. Arestriction map of the DNA insert in homology vector 515-85.1 is givenin FIG. 4. It may be constructed utilizing standard recombinant DNAtechniques (22 and 29), by joining two restriction fragments from thefollowing sources. The first fragment is an approximately 2972 base pairHindIII to BamHI restriction fragment of pSP64 (Promega). The secondfragment is an approximately 3628 base pair HindIII to BclII restrictionsub-fragment of the SPV HindIII restriction fragment M (23).

HOMOLOGY VECTOR 520-17.5. The plasmid 520-17.5 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliβ-galactosidase (lacZ) marker gene flanked by SPV DNA. Upstream of themarker gene is an approximately 2149 base pair fragment of SPV DNA.Downstream of the marker gene is an approximately 1484 base pairfragment of SPV DNA. When this plasmid is used according to theHOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV avirus containing DNA coding for the marker gene will result. Note thatthe β-galactosidase (lacZ) marker gene is under the control of asynthetic early/late pox promoter. A detailed description of the plasmidis given in FIG. 4. It may be constructed utilizing standard recombinantDNA techniques (22 and 30), by joining restriction fragments from thefollowing sources with the synthetic DNA sequences indicated in FIG. 4.The plasmid vector is derived from an approximately 2972 base pairHindIII to BamHI restriction fragment of pSP64 (Promega). Fragment 1 isan approximately 2149 base pair HindIII to AccI restriction sub-fragmentof the SPV HindIII restriction fragment M (23). Fragment 2 is anapproximately 3006 base pair BamHI to PvuII restriction fragment ofplasmid pJF751 (11). Fragment 3 is an approximately 1484 base pair AccIto BglII restriction sub-fragment of the SPV HindIII fragment M (23).

HOMOLOGY VECTOR 538-46.16. The plasmid 538-46.16 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliβ-galactosidase (lacZ) marker gene and the PRV g50 (gpD) gene flanked bySPV DNA. Upstream of the foreign genes is an approximately 2149 basepair fragment of SPV DNA. Downstream of the foreign genes is anapproximately 1484 base pair fragment of SPV DNA. When this plasmid isused according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATINGRECOMBINANT SPV a virus containing DNA coding for the foreign genes willresult. Note that the β-galactosidase (lacZ) marker gene is under thecontrol of a synthetic late pox promoter (LP1) and the g50 (gpD) gene isunder the control of a synthetic early/late pox promoter (EP1LP2). Adetailed description of the plasmid is given in FIG. 5. It may beconstructed utilizing standard recombinant DNA techniques (22 and 30),by joining restriction fragments from the following sources with thesynthetic DNA sequences indicated in FIG. 5. The plasmid vector isderived from an approximately 2972 base pair HindIII to BamHIrestriction fragment of pSP64 (Promega). Fragment 1 is an approximately2149 base pair HindIII to AccI restriction sub-fragment of the SPVHindIII restriction fragment M (23). Fragment 2 is an approximately 3006base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11).Fragment 3 is an approximately 1571 base pair EcoRI to StuI restrictionsub-fragment of the PRV BamHI fragment 7 (21). Note that the EcoRI sitewas introduced into this fragment by PCR cloning. In this procedure theprimers described below were used along with a template consisting of aPRV BamHI #7 fragment subcloned into pSP64. The first primer 87.03(5'-CGCGAATTCGCTCG CAGCGCTATTGGC-3') (SEQ ID NO:41) sits down on the PRVg50 (gpD) sequence (26) at approximately amino acid 3 priming toward the3' end of the gene. The second primer 87.06 (5'-GTAGGAGTGGCTGCTGAAG-3')(SEQ ID NO:42) sits down on the opposite strand at approximately aminoacid 174 priming toward the 5' end of the gene. The PCR product may bedigested with EcoRI and SalI to produce an approximately 509 base pairfragment. The approximately 1049 base pair SalI to StuI sub-fragment ofPRV BamHI #7 may then be ligated to the approximately 509 base pairEcoRI to SalI fragment to generate the approximately 1558 base pairEcoRI to StuI fragment 3. Fragment 4 is an approximately 1484 base pairAccI to BglII restriction sub-fragment of the SPV HindIII fragment M(23).

HOMOLOGY VECTOR 570-91.21. The plasmid 570-91.21 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliB-galactosidase (lacZ) marker gene and the PRV gIII (gpC) gene flankedby SPV DNA. Upstream of the foreign DNA genes is an approximately 1484base pair fragment of SPV DNA. Downstream of the foreign genes is anapproximately 2149 base pair fragment of SPV DNA. When this plasmid isused according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATINGRECOMBINANT SPV, a virus containing DNA coding for the foreign geneswill result. Note that the β-galactosidase (lacZ) marker gene is underthe control of a synthetic late pox promoter (LP1), and the gIII (gpC)gene is under the control of a synthetic early pox promoter (EP2). Adetailed description of the plasmid is given in FIG. 10. It may beconstructed utilizing standard recombinant DNA techniques (22 and 30),by joining restriction fragments from the following sources with thesynthetic DNA sequences indicated in FIG. 10. The plasmid vector isderived from an approximately 2972 base pair HindIII to BamHIrestriction fragment of pSP64 (Promega). Fragment 1 is an approximately1484 base pair BglII to AccI restriction sub-fragment of the SPV HindIIIrestriction fragment M (23). Fragment 2 is an approximately 3002 basepair BamHI to PvuII restriction fragment of plasmid pJF751 (11).Fragment 3 is an approximately 2378 base pair NcoI to NcoI fragment ofplasmid 251-41.A, a subfragment of PRV BamHI #2 and #9. EcoRI linkershave replaced the NcoI and NcoI sites at the ends of this fragment.Fragment 4 is an approximately 2149 base pair AccI to HindIIIrestriction sub-fragment of the SPV HindIII fragment M (23). The AccIsites in fragments 1 and 4 have been converted to PstI sites usingsynthetic DNA linkers.

HOMOLOGY VECTOR 570-91.41. The plasmid 570-91.41 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliB-galactosidase (lacz) marker gene and the PRV gIII (gpC) gene flankedby SPV DNA. Upstream of the foreign DNA genes is an approximately 2149base pair fragment of SPV DNA. Downstream of the foreign genes is anapproximately 1484 base pair fragment of SPV DNA. When this plasmid isused according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATINGRECOMBINANT SPV, a virus containing DNA coding for the foreign geneswill result. Note that the β-galactosidase (lacZ) marker gene is underthe control of a synthetic late pox promoter (LP1), and the gIII (gpC)gene is under the control of a synthetic early late pox promoter(EP1LP2). A detailed description of the plasmid is given in FIG. 11. Itmay be constructed utilizing standard recombinant DNA techniques (22 and30), by joining restriction fragments from the following sources withthe synthetic DNA sequences indicated in FIG. 11. The plasmid vector isderived from an approximately 2972 base pair HindIII to BamHIrestriction fragment of pSP64 (Promega). Fragment 1 is an approximately1484 base pair BglII to AccI restriction sub-fragment of the SPV HindIIIrestriction fragment M (23). Fragment 2 is an approximately 3002 basepair BamHI to PvuII restriction fragment of plasmid pJF751 (11).Fragment 3 is an approximately 2378 base pair NcoI to NcoI fragment ofplasmid 251-41.A, a subfragment of PRV BamHI #2 and #9. EcoRI linkershave replaced the NcoI and NcoI sites at the ends of this fragment.Fragment 4 is an approximately 2149 base pair AccI to HindIIIrestriction sub-fragment of the SPV HindIII fragment M (23). The AccIsites in fragments 1 and 4 have been converted to PstI sites usingsynthetic DNA linkers.

HOMOLOGY VECTOR 570-91.64. The plasmid 570-91.64 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliB-galactosidase (lacZ) marker gene and the PRV gIII (gpC) gene flankedby SPV DNA. Upstream of the foreign DNA genes is an approximately 1484base pair fragment of SPV DNA. Downstream of the foreign genes is anapproximately 2149 base pair fragment of SPV DNA. When this plasmid isused according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATINGRECOMBINANT SPV, a virus containing DNA coding for the foreign geneswill result. Note that the β-galactosidase (lacZ) marker gene is underthe control of a synthetic late pox promoter (LP1), and the gIII (gpC)gene is under the control of a synthetic late early pox promoter(LP2EP2). A detailed description of the plasmid is given in FIG. 12. Itmay be constructed utilizing standard recombinant DNA techniques (22 and30), by joining restriction fragments from the following sources withthe synthetic DNA sequences indicated in FIG. 12. The plasmid vector isderived from an approximately 2972 base pair HindIII to BamHIrestriction fragment of pSP64 (Promega). Fragment 1 is an approximately1484 base pair BglII to AccI restriction sub-fragment of the SPV HindIIIrestriction fragment M (23). Fragment 2 is an approximately 3002 basepair BamHI to PvuII restriction fragment of plasmid pJF751 (11).Fragment 3 is an approximately 2378 base pair NcoI to NcoI fragment ofplasmid 251-41.A, a subfragment of PRV BamHI #2 and #9. EcoRI linkershave replaced the NcoI and NcoI sites at the ends of this fragment.Fragment 4 is an approximately 2149 base pair AccI to HindIIIrestriction sub-fragment of the SPV HindIII fragment M (23). The AccIsites in fragments 1 and 4 have been converted to PstI sites usingsynthetic DNA linkers.

HOMOLOGY VECTOR 538-46.26. The plasmid 538-46.26 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E.coliβ-galactosidase (lacZ) marker gene and the Newcastle Disease Virus (NDV)hemagglutinin-Neuraminidase (HN) gene flanked by SPV DNA. Upstream ofthe foreign genes is an approximately 2149 base pair fragment of SPVDNA. Downstream of the foreign genes is an approximately 1484 base pairfragment of SPV DNA. When this plasmid is used according to theHOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV avirus containing DNA coding for the foreign genes will result. Note thatthe β-galactosidase (lacZ) marker gene is under the control of asynthetic late pox promoter (LP1) and the HN gene is under the controlof a synthetic early/late pox promoter (EP1LP2). A detailed descriptionof the plasmid is given in FIG. 8. It may be constructed utilizingstandard recombinant DNA techniques (22 and 30), by joining restrictionfragments from the following sources with the synthetic DNA sequencesindicated in FIG. 8. The plasmid vector is derived from an approximately2972 base pair HindIII to BamHI restriction fragment of pSP64 (Promega).Fragment 1 is an approximately 2149 base pair HindIII to AccIrestriction sub-fragment of the SPV HindIII restriction fragment M (23).Fragment 2 is an approximately 1810 base pair AvaII to NaeI restrictionfragment of a NDV HN cDNA clone. The sequence of the HN cDNA clone isgiven in FIG. 7. The cDNA clone was generated from the Bi strain of NDVusing standard cDNA cloning techniques (14). Fragment 3 is anapproximately 3006 base pair BamHI to PvuII restriction fragment ofplasmid pJF751 (11). Fragment 4 is an approximately 1484 base pair AccIto BglII restriction sub-fragment of the SPV HindIII fragment M (23).

HOMOLOGY VECTOR 599-65.25. The plasmid 599-65.25 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliB-galactosidase (lacZ) marker gene and the ILT gpG gene flanked by SPVDNA. Upstream of the foreign genes is an approximately 1484 base pairfragment of SPV DNA. Downstream of the foreign genes is an approximately2149 base pair fragment of SPV DNA. When the plasmid is used accordingto the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANTSPV, a virus containing DNA coding for the foreign genes will result.Note that the B-galactosidase (lacZ) marker gene is under the control ofa synthetic late pox promoter (LP1), and the ILT gpG gene is under thecontrol of a synthetic early/late pox promoter (EP1LP2). A detaileddescription of the plasmid is given in FIG. 13. It may be constructedutilizing standard recombinant DNA techniques (22, 30), by joiningrestriction fragments from the following sources with the synthetic DNAsequences indicated in FIG. 13. The plasmid vector is derived from anapproximately 2972 base pair HindIII to BamHI restriction fragment ofpSP64 (Promega). Fragment 1 is an approximately 1484 base pair BglII toAccI restriction sub-fragment of the SPV HindIII restriction fragment M(23). Fragment 2 is an approximately 1073 base pair EcoRI to MboIfragment. Note that the EcoRI site was introduced by PCR cloning. Inthis procedure, the primers described below were used with a templateconsisting of a 2.6 kb Sst I to Asp718I subfragment of a 5.1 kbAsp718Ifragment of ILT virus genome. The first primer 91.13(5'-CCGAATTCCGGCTTCAGTAACATAGGATCG-3') (SEQ ID NO: 81) sits down on theILT gpG sequence at amino acid 2. It adds an additional asparagineresidue between amino acids 1 and 2 and also introduces an EcoRIrestriction site. The second primer 91.14 (5'-GTACCCATACTGGTCGTGGC-3')(SEQ ID NO: 82) sits down on the opposite strand at approximately aminoacid 196 priming toward the 5' end of the gene. The PCR product isdigested with EcoRI and BamHI to produce an approximately 454 base pairfragment. The approximately 485 base pair MboI sub-fragment of ILTAsp718I (5.1 kb) fragment is ligated to the approximately 454 base pairEcoRI to BamHI fragment to generate fragment 2 from EcoRI to MboI whichis approximately 939 base pairs (293 amino acids) in length. Fragment 3is an approximately 3002 base pair BamHI to PvuII restriction fragmentof plasmid pJF751 (11). Fragment 4 is an approximately 2149 base pairAccI to HindIII subfragment of the SPV HindIII fragment M. The AccIsites of fragments 1 and 4 have been converted to PstI sites usingsynthetic DNA linkers.

HOMOLOGY VECTOR 624-20.1C. The plasmid 624-20.lC was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliB-galactosidase (lacZ) marker gene and the ILT gpI gene flanked by SPVDNA. Upstream of the foreign genes is an approximately 1484 base pairfragment of SPV DNA. Downstream of the foreign genes is an approximately2149 base pair fragment of SPV DNA. When the plasmid is used accordingto the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANTSPV, a virus containing DNA coding for the foreign genes will result.Note that the B-galactosidase (lacZ) marker gene is under the control ofa synthetic late pox promoter (LP1), and the ILT gpI gene is under thecontrol of a synthetic late/early pox promoter (LP2EP2). A detaileddescription of the plasmid is given in FIG. 14. It may be constructedutilizing standard recombinant DNA techniques (22, 30), by joiningrestriction fragments from the following sources with the synthetic DNAsequences indicated in FIG. 14. The plasmid vector is derived from anapproximately 2972 base pair HindIII to BamHI restriction fragment ofpSP64 (Promega). Fragment 1 is an approximately 1484 base pair Bgl II toAccI restriction sub-fragment of the SPV HindIII restriction fragment M(23). Fragment 2 is an approximately 1090 base pair fragment with EcoRIand BamHI restriction sites at the ends synthesized by PCR cloning andcontaining the entire amino acid coding sequence of the ILT gpI gene.The ILT gpI gene was synthesized in two separate PCR reactions. In thisprocedure, the primers described below were used with a templateconsisting the 8.0 kb ILT Asp 7181 fragment. The first primer 103.6(5'-CCGGAATTCGCTACTT GGAACTCTGG-3') (SEQ ID NO 83) sits down on the ILTgpI sequence at amino acid number 2 and introduces an EcoRI site at the5' end of the ILT gpI gene, The second primer 103.3(5'-CATTGTCCCGAGACGGACAG-3') (SEQ ID NO. 84) sits down on the ILT gpIsequence at approximately amino acid 269 on the opposite strand toprimer 103.6 and primes toward the 5' end of the gene. The PCR productwas digested with EcoRI and BglI (BglI is located approximately at aminoacid 209 which is 179 base pairs 5' to primer 2) to yield a fragment 625base pairs in length corresponding to the 5' end of the ILT gpI gene.The third primer 103.4 (5'-CGCGATCCAACTATCGGTG-3') (SEQ ID NO. 85) sitsdown on the ILT gpI gene at approximately amino acid 153 priming towardthe 3' end of the gene. The fourth primer 103.5 (5'GCGGATCCACATTCAGACTTAATCAC-3') (SEQ ID NO. 86) sits down at the 3' end of the ILT gpIgene 14 base pairs beyond the UGA stop codon, introducing a BamHIrestriction site and priming toward the 5' end of the gene. The PCRproduct is digested with Bgl I (at amino acid 209) and BamHI to yield afragment 476 base pairs in length corresponding to the 3' end of the ILTgpI gene. Fragment 2 consists of the products of the two PCR reactionsligated together to yield an ILT gpI gene which is a EcoRI to BamHIfragment approximately 1101 base pairs (361 amino acids) in length.Fragment 3 is an approximately 3002 base pair BamHI to PvuII restrictionfragment of plasmid pJF751 (11). Fragment 4 is an approximately 2149base pair AccI to HindIII subfragment of the SPV HindIII fragment M. TheAccI sites in fragments 1 and 4 were converted to unique NotI sitesusing NotI linkers.

HOMOLOGY VECTOR 614-83.18. The plasmid 614-83.18 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliB-galactosidase (lacZ) marker gene and the IBR gpG gene flanked by SPVDNA. Upstream of the foreign genes is an approximately 1484 base pairfragment of SPV DNA. Downstream of the foreign genes is an approximately2149 base pair fragment of SPV DNA. When the plasmid is used accordingto the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANTSPV, a virus containing DNA coding for the foreign genes will result.Note that the B-galactosidase (lacZ) marker gene is under the control ofa synthetic late pox promoter (LP1), and the IBR gG gene is under thecontrol of a synthetic late/early pox promoter (LP2EP2). A detaileddescription of the plasmid is given in FIG. 15. It may be constructedutilizing standard recombinant DNA techniques (22, 30), by joiningrestriction fragments from the following sources with the synthetic DNAsequences indicated in FIG. 15. The plasmid vector is derived from anapproximately 2972 base pair HindIII to BamHI restriction fragment ofpSP64 (Promega). Fragment 1 is an approximately 1484 base pair BglII toAccI restriction sub-fragment of the SPV HindIII restriction fragment M(23). Fragment 2 is an approximately 1085 base pair fragment synthesizedby PCR cloning with EcoRI and BamHI restriction sites at the ends andcontaining the amino acid coding sequence from amino acids 2 to 362 ofthe IBR gpG gene. In the PCR cloning procedure, the primers describedbelow were used with a template consisting of the IBR-000 virus (Cooperstrain). The first primer 106.9 (5'-ATGAATTCCCCTGCCGCCCGGACCGGCACC-3')(SEQ ID NO. 87) sits down on the IBR gpG sequence at amino acid number 1and introduces an EcoRI site at the 5' end of the IBR gpG gene and twoadditional amino acids between amino acids 1 and 2. The second primer106.8 (5'-CATGGATCCCGCTCGAGGCGAGCGGGCTCC-3') (SEQ ID NO. 88) sits downon the IBR gpG sequence at approximately amino acid 362 on the Oppositestrand to primer 1 and primes synthesis toward the 5' end of the IBR gpGgene. Fragment 2 was generated by digesting the PCR product with EcoRIand BamHI to yield a fragment 1085 base pairs in length corresponding tothe amino terminal 362 amino acids (approximately 80%) of the IBR gpGgene. Fragment 3 is an approximately 3002 base pair BamHI to PvuIIrestriction fragment of plasmid pJF751 (11). Fragment 4 is anapproximately 2149 base pair AccI to HindIII subfragment of the SPVHindIII fragment M. The AccI sites in fragments 1 and 4 were convertedto unique NotI sites using NotI linkers.

Homology Vector for Constructing S-SPV-019 (LacZ/IBR gpE HomologyVector)

This lacZ/IBR gpE homology vector is used to insert foreign DNA intoSPV. It incorporates an E. coli B-galactosidase (lacZ) marker gene andthe IBR gpE gene flanked by SPV DNA. When this plasmid is used accordingto the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPVa virus containing DNA coding for the foreign genes will result. Notethat the B-galactosidase (lacZ) marker gene is under the control of asynthetic late pox promoter and the gpE gene is under the control of asynthetic late/early pox promoter. The homology vector may beconstructed utilizing standard recombinant DNA techniques (22 and 30),by joining restriction fragments from the following sources with theappropriate synthetic DNA sequences. The plasmid vector is derived froman approximately 2972 base pair HindIII to BamHI restriction fragment ofpSP64 (Promega). The upstream SPV homology is an approximately 1146 basepair BglIII to AccI restriction sub-fragment of the SPV HindIII fragmentM (23). The IBR gE gene is an approximately 1888 base pair fragmentsynthesized by PCR cloning with EcoRI and BamHI ends. In the PCR cloningprocedure, the primers described below were used with a templateconsisting of the IBR-000 VIRUS (Cooper strain). The first primer4/93.17DR (5'-CTGGTTCGGCCCAGAATTCTATGGGTCTCGCGCGGCTCGTGG-3' (SEQ ID NO.89) sits down on the IBR gpE gene at amino acid number 1 and introducesan EcoRI site at the 5' end of the IBR gpE gene and adds two additionalamino acids at the amino terminus of the protein. The second primer4/93.18DR (5'-CTCGCTCGCCCAGGATCCCTAGCGGAGGATGGACTTGAGTCG-3') (SEQ ID NO.90) sits down on the IBR gpE sequence at approximately amino acid 648 onthe opposite strand to primer 1 and primes synthesis toward the 5' endof the IBR gpE gene. The lacZ promoter and marker gene is identical tothe one used in plasmid 520-17.5. The downstream SPV homology is anapproximately 2156 base pair AccI to HindIII restriction sub-fragment ofthe SPV HindIII restriction fragment M (23). The AccI site in the SPVhomology vector is converted to a unique XbaI site.

Homology Vector for Constructing S-SPV-018 (LacZ/PRV gpE HomologyVector)

This homology vector is constructed for the purpose of inserting foreignDNA into SPV. It incorporates an E. coli B-galactosidase (lacZ) markergene and the PRV gpE gene flanked by SPV DNA. Upstream of the foreigngenes is an approximately 1484 base pair fragment of SPV DNA. Downstreamof the foreign genes is an approximately 2149 base pair fragment of SPVDNA. When the plasmid is used according to the HOMOLOGOUS RECOMBINATIONPROCEDURE FOR GENERATING RECOMBINANT SPV, a virus containing the DNAcoding for the foreign genes results. Note that the B- galactosidase(lacz) marker gene is under the control of a synthetic late pox promoter(LP1), and the PRV gpE gene is under the control of a syntheticearly/late pox promoter (EP1LP2). The homology vector is constructedutilizing standard recombinant DNA techniques (22, 30), by joiningrestriction fragments from the following sources with synthetic DNAsequences. The plasmid vector is derived from an approximately 2972 basepair HindIII to BamHI restriction fragment pSP64 (Promega). Fragment 1is an approximately 1484 base pair BglII to AccI restrictionsub-fragment of the SPV HindIII restriction fragment M (23). Fragment 2is the lacZ promoter and marker gene which is identical to the one usedin plasmid 520-17.5. Fragment 3 is an approximately 2484 base pair DraIto MluI sub-fragment of PRV derived from the PRV BamHI #7 DNA fragment.The DraI site is converted to an EcoRI site through the use of asynthetic DNA linker. The DraI site sits 45 base pairs upstream of thenatural gpE start codon and extends the open reading frame at the aminoterminus of the protein for 15 amino acids. The synthetic poxpromoter/EcoRI DNA linker contributes another 4 amino acids. Therefore,the engineered gpE gene contains 19 additional amino acids fused to theamino terminus of gpE. The nineteen amino acids areMet-Asn-Ser-Gly-Asn-Leu-Gly-Thr-Pro-Ala-Ser-Leu-Ala-His-Thr-Gly-Val-Glu-Thr.Fragment 4 is an approximately 2149 base pair AccI to HindIIIsub-fragment of the SPV HindIII fragment M (23). The AccI sites offragments 1 and 4 are converted to PstI sites using synthetic DNAlinkers.

HOMOLOGY VECTOR 520-90.15. The plasmid 520-90.15 was constructed for thepurpose of inserting foreign DNA into SPV. It contains a unique NdeIrestriction enzyme site into which foreign DNA may be inserted. When aplasmid, containing a foreign DNA insert at the NdeI site, is usedaccording to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATINGRECOMBINANT SPV a virus containing the foreign DNA will result. Plasmid520-90.15 was constructed utilizing standard recombinant DNA techniques(22 and 30), by joining two restriction fragments from the followingsources. The first fragment is an approximately 2972 base pair HindIIIto BamHI restriction fragment of pSP64 (Promega). The second fragment isan approximately 1700 base pair HindIII to BamHI restriction subfragmentof the SPV HindIII restriction fragment G (23).

HOMOLOGY VECTOR 708-78.9. The plasmid 708-78.9 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliβ-galactosidase (lacZ) marker gene and the infectious bovinerhinotracheitis virus (IBRV) gE gene flanked by SPV DNA. Upstream of theforeign genes is an approximately 1484 base pair fragment of SPV DNA.Downstream of the foreign genes is an approximately 2149 base pairfragment of SPV DNA. When the plasmid is used is according to theHOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV, avirus containing DNA coding for the foreign genes will result. Note thatthe β-galactosidase (lacZ) marker gene is under the control of asynthetic late pox promoter (LP1), and the IBRV gE gene is under thecontrol of a synthetic late/early pox promoter (LP2EP2). It may beconstructed utilizing standard recombinant DNA techniques (22, 30), byjoining restriction fragments from the following sources. The plasmidvector is derived from an approximately 2972 base pair HindIII to BamHIrestriction fragment of pSP64 (Promega). Fragment 1 is an approximately1484 base pair Bgl II to AccI restriction sub-fragment of the SPVHindIII restriction fragment M (23). Fragment 2 is an approximately 475base pair fragment with EcoRI and BamHI restriction sites at the ends.The EcoRI and BamHI sites are synthesized by PCR cloning. The PCRproduct contains the entire amino acid coding sequence of the IBRV gEgene. In the PCR cloning procedure, the primers described below wereused with a template consisting of the IBR-000 virus (Cooper strain)(44). The first primer 2/94.5DR(5'-CTGGTTCGGCCCAGAATTCGATGCAACCCACCGCGCCGCCCCG-3') (SEQ ID NO. 116)sits down on the IBR gpE gene at amino acid number 1 and introduces anEcoRI site at the 5' end of the IBRV gE gene and adds two additionalamino acids at the amino terminus of the protein. The second primer4/93.18DR (5'-CTCGCTCGCCCAGGATCCCTAGCGGAGGATGGACTTGAGTCG-3,) (SEQ ID NO.117) sits down on the IBRV gE sequence (44) at approximately amino acid648 on the opposite strand to the first primer and primes synthesistoward the 5' end of the IBRV gE gene. The PCR product was digested withEcoRI and BamHI to yield a fragment approximately 1950 base pairs inlength corresponding to the IBRV gE gene. Fragment 3 is an approximately3002 base pair BamHI to PvuII restriction fragment of plasmid pJF751(11). Fragment 4 is an approximately 2149 base pair AccI to HindIIIsubfragment of the SPV HindIII fragment M. The AccI sites in fragments 1and 4 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 723-59A9.22. The plasmid 723-59A9.22 was used to insertforeign DNA into SPV. It incorporates an E. coli β-galactosidase (lacZ)marker gene and the equine influenza virus NA PR/56 gene flanked by SPVDNA. When this plasmid was used according to the HOMOLOGOUSRECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV a viruscontaining DNA coding for the foreign genes results. Note that theβ-galactosidase (lacZ) marker gene is under the control of a syntheticlate pox promoter (LP1) and the EIV PR/56 NA gene is under the controlof a synthetic late/early pox promoter (LP2EP2). A detailed descriptionof the plasmid is given in FIGS. 18A, 18B, 18C and 18D. The homologyvector was constructed utilizing standard recombinant DNA techniques (22and 30), by joining restriction fragments from the following sourceswith the appropriate synthetic DNA sequences. The plasmid vector isderived from an approximately 2972 base pair HindIII to BamHIrestriction fragment of pSP64 (Promega). Fragment 1 is an approximately1484 base pair BglII to AccI restriction sub-fragment of the SPV HindIIIfragment M (23). Fragment 2 is the NA gene coding region from the equineInfluenza A/Prague/56 (serotype 1 (N7) virus) cloned as an approximately1450 base pair BamHI fragment-utilizing the following primers5'-GGGATCCATGAATCCTAATCAAAAACTCTTT-3' (SEQ ID NO: 118) for cDNA primingand combined with 5'-GGGATCCTTACGAAAAGTATTTAATTTGTGC-3' (SEQ ID NO: 119)for PCR. (see CLONING OF EQUINE INFLUENZA VIRUS HEMAGGLUTININ ANDNEURAMINIDASE GENES). Fragment 3 is an approximately 3010 base pairBamHI to PvuII restriction fragment of plasmid pJF751 (11). Fragment 4is an approximately 2149 base pair AccI to HindIII restrictionsub-fragment of the SPV HindIII restriction fragment M (23). The AccIsite in the SPV homology vector is converted to a unique NotI site.

HOMOLOGY VECTOR 727-54.60. The plasmid 727-54.60 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliβ-galactosidase (lacz) marker gene and the pseudorabies virus (PRV) gII(gpB) gene flanked by SPV DNA. Upstream of the foreign genes is anapproximately 1484 base pair fragment of SPV DNA. Downstream of theforeign genes is an approximately 2149 base pair fragment of SPV DNA.When the plasmid is used according to the HOMOLOGOUS RECOMBINATIONPROCEDURE FOR GENERATING RECOMBINANT SPV, a virus containing DNA codingfor the foreign genes will result. Note that the β-galactosidase (lacZ)marker gene is under the control of a synthetic late pox promoter (LP1),and the PRV gB gene is under the control of a synthetic late/early poxpromoter (LP2EP2). A detailed description of the plasmid is given inFIGS. 19A, 19B, 19C, and 19D. It may be constructed utilizing standardrecombinant DNA techniques (22, 30), by joining restriction fragmentsfrom the following sources with the synthetic DNA sequences indicated inFIGS. 19A to 19D. The plasmid vector is derived from an approximately2972 base pair HindIII to BamHI restriction fragment of pSP64 (Promega).Fragment 1 is an approximately 1484 base pair BglII to AccI restrictionsub-fragment of the SPV HindIII restriction fragment M (23). Fragment 2is an approximately 3500 base pair fragment which contains the codingsequence for the PRV gB gene within the KpnI C fragment of genomic PRVDNA(21). Fragment 2 contains an approximately 53 base pair syntheticfragment containing the amino terminus of the PRV gB gene, anapproximately 78 base pair SmaI to Nhe I fragment from the PRV KpnI Cgenomic fragment, and an approximately 3370 base pair NheI to EcoRIfragment from the PRV KpnI C genomic fragment (21). Fragment 3 is anapproximately 3010 base pair BamHI to PvuII restriction fragment ofplasmid pJF751 (11). Fragment 4 is an approximately 2149 base pair AccIto HindIII subfragment of the SPV HindIII fragment M. The AccI sites infragments 1 and 4 were-converted to unique NotI sites using NotIlinkers.

HOMOLOGY VECTOR 727-67.18. The plasmid 727-67.18 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliβ-galactosidase (lacZ) marker gene and the hepatitis B virus coreantigen gene flanked by SPV DNA. Upstream of the foreign genes is anapproximately 1484 base pair fragment of SPV DNA. Downstream of theforeign genes is an approximately 2149 base pair fragment of SPV DNA.When the plasmid is used according to the HOMOLOGOUS RECOMBINATIONPROCEDURE FOR GENERATING RECOMBINANT SPV, a virus containing DNA codingfor the foreign genes will result. Note that the β-galactosidase (lacZ)marker gene is under the control of a synthetic late pox promoter (LP1),and the hepatitis B core antigen gene is under the control of asynthetic early/late pox promoter (EP1LP2). A detailed description ofthe plasmid is given in FIGS. 20A, 20B, 20C and 20D. It may beconstructed utilizing standard recombinant DNA techniques (22, 30), byjoining restriction fragments from the following sources with thesynthetic DNA sequences indicated in FIGS. 20A to 20D. The plasmidvector is derived from an approximately 2972 base pair HindIII to BamHIrestriction fragment of pSP64 (Promega). Fragment 1 is an approximately1484 base pair BglII to AccI restriction sub-fragment of the SPV HindIIIrestriction fragment M (23). Fragment 2 is an approximately 3002 basepair BamHI to PvuII restriction fragment of plasmid pJF751 (11).Fragment 3 is an approximately 589 base pair fragment with BamHI andEcoRI restriction sites at the ends. This fragment contains thehepatitis B core antigen coding sequences (amino acids 25-212) (Ref. 45,50). Fragment 4 is an approximately 2149 base pair AccI to HindIIIsubfragment of the SPV HindIII fragment M. The AccI sites in fragments 1and 4 were converted to unique NotI sites using NotI linkers.

CLONING OF EQUINE INFLUENZA VIRUS HEMAGGLUTININ AND NEURAMINIDASE GENES.The equine influenza virus hemagglutinin (HA) and Neuraminidase (NA)genes may be cloned essentially as described by Katz et al. (42) for theHA gene of human influenza virus. Viral RNA prepared from virus grown inMDBK cells (for Influenza A/equine/Alaska/91 and InfluenzaA/equine/Miami/63) and MDCK cells (for Influenza A/equine/Prague/56 andInfluenza A/equine/Kentucky/81) is first converted to cDNA utilizing anoligo nucleotide primer specific for the target gene. The cDNA is thenused as a template for PCR cloning (51) of the targeted gene region. ThePCR primers are designed to incorporate restriction sites which permitthe cloning of the amplified coding regions into vectors containing theappropriate signals for expression in EHV. One pair of oligo nucleotideprimers will be required for each coding region. The HA gene codingregions from the serotype 2 (H3) viruses (Influenza A/equine/Miami/63,Influenza A/equine/Kentucky/81, and Influenza A/equine/Alaska/91) wouldbe cloned utilizing the following primers5'-GGAGGCCTTCATGACAGACAACCATTATTTTGATACTACTGA-3' (SEQ ID NO: 120) forcDNA priming and combined with5'-GAAGGCCTTCTCAAATGCAAATGTTGCATCTGATGTTGCC-3' (SEQ ID NO: 121) for PCR.The HA gene coding region from the serotype 1 (H7) virus (InfluenzaA/equine/Prague/56) would be cloned utilizing the following primers5'-GGGATCCATGAACACTCAAATTCTAATATTAG-3' (SEQ ID NO: 122) for cDNA primingand combined with 5'-GGGATCCTTATATACAAATAGTGCACCGCA-3' (SEQ ID NO: 123)for PCR. The NA gene coding regions from the serotype 2 (N8) viruses(Influenza A/equine/Miami/63, Influenza A/equine/Kentucky/81, andInfluenza A/equine/Alaska/91) would be cloned utilizing the followingprimers 5'-GGGTCGACATGAATCCAAATCAAAAGATAA-3' (SEQ ID NO: 124) for cDNApriming and combined with 5'-GGGTCGACTTACATCTTATCGATGTCAAA-3' (SEQ IDNO: 125) for PCR. The NA gene coding region from the serotype 1 (N7)virus (Influenza/A/equine/Prague/56) would be cloned utilizing thefollowing primers 5'-GGGATCCATGAATCCTAATCAAAAACTCTTT-3' (SEQ ID NO: 118)for cDNA priming and combined with 5'-GGGATCCTTACGAAAAGTATTTAATTTGTGC-3'(SEQ ID NO: 119) for PCR. Note that this general strategy may be used toclone the coding regions of HA and NA genes from other strains of equineinfluenza A virus. The EIV HA or NA genes are cloned as a blunt endedSalI or BamHI fragment into a blunt ended EcoRI site behind the LP2EP2promoter of the SPV homology vector.

HOMOLOGY VECTOR 732-18.4. The plasmid 732-18.4 was used to insertforeign DNA into SPV. It incorporates an E. coli β-galactosidase (lacZ)marker gene and the equine influenza virus AK/91 NA gene flanked by SPVDNA. When this plasmic was used according to the HOMOLOGOUSRECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV a viruscontaining DNA coding for the foreign genes results. Note that theβ-galactosidase (lacZ) marker gene is under the control of a syntheticlate pox promoter (LP1) and the EIV AK/91 NA gene is under the controlof a synthetic late/early pox promoter (LP2EP2). A detail description ofthe plasmid is given in FIGS. 21A, 21B, 21C and 21D. The homology vectorwas constructed utilizing standard recombinant DNA techniques (22 and30), by joining restriction fragments from the following sources withthe appropriate synthetic DNA sequences. The plasmid vector is derivedfrom an approximately 2972 base pair HindIII to BamHI restrictionfragment of pSP64 (Promega). Fragment 1 is an approximately 1484 basepair BglII to AccI restriction sub-fragment of the SPV HindIII fragmentM (23). Fragment 2 is the NA gene coding region from the equineInfluenza A/Alaska/91 (serotype 2 (N8) virus) cloned as an approximately1450 base pair SalI fragment utilizing the following primers5'-GGGTCGACATGAATCCAAATCAAAAGATAA-3' (SEQ ID NO: 124) for cDNA primingand combined with 5'-GGGTCGACTTACATCTTATCGATGTCAAA-3' (SEQ ID NO: 125)for PCR (see CLONING OF EQUINE INFLUENZA VIRUS HEMAGGLUTININ ANDNEURAMINIDASE GENES). Fragment 3 is an approximately 3010 base pairBamHI to PvuII restriction fragment of plasmid pJF751 (11). Fragment 4is an approximately 2149 base pair AccI to HindIII restrictionsub-fragment of the SPV HindIII restriction fragment M (23). The AccIsite in the SPV homology vector is converted to a unique NotI site.

HOMOLOGY VECTOR 741-80.3 The plasmid 741-80.3 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliβ-galactosidase (lacZ) marker gene flanked by SPV DNA. Upstream of theforeign genes is an approximately 1484 base pair fragment of SPV DNA.Downstream of the foreign genes is an approximately 2149 base pairfragment of SPV DNA. When the plasmid is used according to theHOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV, avirus containing DNA coding for the foreign genes will result. Note thatthe β-galactosidase (lacZ) marker gene is under the control of a humancytomegalovirus immediate early (HCMV IE) promoter. A detaileddescription of the plasmid is given in FIGS. 22A, 22B and 22C. It may beconstructed utilizing standard recombinant DNA techniques (22, 30), byjoining restriction fragments from the following sources with thesynthetic DNA sequences indicated in FIGS. 22A to 22C. The plasmidvector is derived from an approximately 2972 base pair HindIII to BamHIrestriction fragment of pSP64 (Promega). Fragment 1 is an approximately1484 base pair BglII to AccI restriction sub-fragment of the SPV HindIIIrestriction fragment M (23) Fragment 2 is a 1154 base pair PstI to AvaIIfragment derived from a HCMV 2.1 kb PstI fragment containing the HCMV IEpromoter (46). Fragment 3 is a 3010 base pair BamHI to PvuII fragmentderived from plasmid pJF751 (49) containing the E. coli lacZ gene.Fragment 4 is an approximately 750 base pair NdeI to SalI fragmentderived from PRV BamHI #7 which contains the carboxy-terminal 19 aminoacids and the polyadenylation signal of the PRV gX gene. Fragment 5 isan approximately 2149 base pair AccI to HindIII subfragment of the SPVHindIII fragment M. The AccI sites in fragments 1 and 5 were convertedto unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 741-84.14. The plasmid 741-84.14 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliβ-galactosidase (lacZ) marker gene and the human interleukin-2 (IL-2)gene flanked by SPV DNA. Upstream of the foreign genes is anapproximately 1484 base pair fragment of SPV DNA. Downstream of theforeign genes is an approximately 2149 base pair fragment of SPV DNA.When the plasmid is used according to the HOMOLOGOUS RECOMBINATIONPROCEDURE FOR GENERATING RECOMBINANT SPV, a virus containing DNA codingfor the foreign genes will result. Note that the β-galactosidase (lacZ)marker gene is under the control of a synthetic late pox promoter (LP1),and the human IL-2 gene is under the control of a synthetic late/earlypox promoter (LP2EP2). The coding sequence for the human IL-2 protein isfused at the amino terminus to the PRV gX signal sequence for membranetransport. A detailed description of the plasmid is given in FIGS. 23A,23B, 23C, and 23D. It may be constructed utilizing standard recombinantDNA techniques (22, 30), by joining restriction fragments from thefollowing sources with the synthetic DNA sequences indicated in FIGS.23A to 23D. The plasmid vector is derived from an approximately 2972base pair HindIII to BamHI restriction fragment of pSP64 (Promega).Fragment 1 is an approximately 1484 base pair BglII to AccI restrictionsub-fragment of the SPV HindIII restriction fragment M (23). Fragment 2is an approximately 475 base pair fragment with EcoRI and BglIIrestriction sites at the ends. The EcoRI site is synthesized by PCRcloning and the BglII site is at the 3' end of the human IL-2 cDNA (43,47). The PCR product contains the entire amino acid coding sequence ofthe PRV gX signal sequence-human IL-2 gene. In this procedure, theprimers described below were used with a template consisting of the cDNAfor PRV gX signal sequence-human IL-2 (43). The first primer 5/94.23(5'-CTCGAATTCGAAGTGGGCAACGTGGATCCTCGC-3') (SEQ ID NO 126) sits down onthe PRV gX signal sequence at amino acid number 1 and introduces anEcoRI site at the 5' end of the gene. The second primer 5/94.24(5'-CAGTTAGCCTCCCCCATCTCCCCA-3') (SEQ ID NO. 127) sits down on the humanIL-2 gene sequence within the 3' untranslated region on the oppositestrand to primer 5/94.23 and primes toward the 5 end of the gene. ThePCR product was digested with EcoRI and BglII (BglII is locatedapproximately 3 nucleotides beyond the stop codon for the human IL-2gene, to yield a fragment 475 base pairs in length corresponding to thePRV gX signal sequence-human IL-2 gene. Fragment 3 is an approximately3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751(11). Fragment 4 is an approximately 2149 base pair AccI to HindIIIsubfragment of the SPV HindIII fragment M. The AccI sites in fragments 1and 4 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 744-34. The plasmid 744-34 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliβ-galactosidase (lacZ) marker gene and the equine herpesvirus type 1 gBgene flanked by SPV DNA. Upstream of the foreign genes is anapproximately 1484 base pair fragment of SPV DNA. Downstream of theforeign genes is an approximately 2149 base pair fragment of SPV DNA.When the plasmid is used according to the HOMOLOGOUS RECOMBINATIONPROCEDURE FOR GENERATING RECOMBINANT SPV, a virus containing DNA codingfor the foreign genes will result. Note that the β-galactosidase (lacZ)marker gene is under the control of a synthetic late pox promoter (LP1),and the EHV-1 gB gene is under the control of a synthetic late/early poxpromoter (LP2EP2). A detailed description of the plasmid is given inFIGS. 24A, 24B, 24C and 24D. It may be constructed utilizing standardrecombinant DNA techniques (22, 30), by joining restriction fragmentsfrom the following sources with the synthetic DNA sequences indicated inFIGS. 24A to 24D The plasmid vector is derived from an approximately2972 base pair HindIII to BamHI restriction fragment of pSP64 (Promega).Fragment 1 is an approximately 1484 base pair Bgl II to AccI restrictionsub-fragment of the SPV HindIII restriction fragment M (23). Fragment 2is an approximately 2941 base pair fragment with EcoRI and PmeIrestriction sites at the ends. Fragment 2 is an approximately 2941 basepair EcoRI to PmeI fragment. Fragment 2 was synthesized as anapproximately 429 base pair PCR fragment at the 5' end of the genehaving a synthetic EcoRI site and a natural BamHI site within the BamHI"a" fragment of EHV-1 genomic DNA and an approximately 2512 base pairrestriction fragment at the 3' end of the gene from BamHI to PmeI withinthe BamHI "i" fragment of EHV-1 genomic DNA (48). In the procedure toproduce the 5' end PCR fragment, the primers described below were usedwith a template consisting of the EHV-1 BamHI "a" and "i" fragments. Thefirst primer 5/94.3 (5'-CGGAATTCCTCTGGTTGCCGT-3') (SEQ ID NO 128) sitsdown on the EHV-1 gB sequence at amino acid number 2 and introduces anEcoRI site at the 5' end of the EHV-1 gB gene and an ATG start codon.The second primer 5/94.4 (5'-GACGGTGGATCCGGTAGGCGGT-3') (SEQ ID NO. 129)sits down on the EHV-1 gB sequence at approximately amino acid 144 onthe opposite strand to primer 5/94.3 and primes toward the 5' end of thegene. The PCR product was digested with EcoRI and BamHI to yield afragment 429 base pairs in length corresponding to the 5' end of theEHV-1 gB gene. Fragment 2 consists of the products of the PCR reaction(EcoRI to BamHI) and the restriction fragment (BamHI to PmeI) ligatedtogether to yield an EHV-1 gB gene which is an EcoRI to PmeI fragmentapproximately 2941 base pairs (979 amino acids) in length. Fragment 3 isan approximately 3010 base pair BamHI to PvuII restriction fragment ofplasmid pJF751 (11). Fragment 4 is an approximately 2149 base pair AccIto HindIII subfragment of the SPV HindIII fragment M. The AccI sites infragments 1 and 4 were converted to unique NotI sites using NotIlinkers.

HOMOLOGY VECTOR 744-38. The plasmid 744-38 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliβ-galactosidase (lacZ) marker gene and the equine herpesvirus type 1 gDgene flanked by SPV DNA. Upstream of the foreign genes is anapproximately 1484 base pair fragment of SPV DNA. Downstream of theforeign genes is an approximately 2149 base pair fragment of SPV DNA.When the plasmid is used according to the HOMOLOGOUS RECOMBINATIONPROCEDURE FOR GENERATING RECOMBINANT SPV, a virus containing DNA codingfor the foreign genes will result. Note that the β-galactosidase (lacZ)marker gene is under the control of a synthetic late pox promoter (LP1),and the EHV-1 gD gene is under the control of a synthetic late/early poxpromoter (LP2EP2). A detailed description of the plasmid is given inFIGS. 25A, 25B, 25C and 25D. It may be constructed utilizing standardrecombinant DNA techniques (22, 30), by joining restriction fragmentsfrom the following sources with the synthetic DNA sequences indicated inFIGS. 25A to 25D. The plasmid vector is derived from an approximately2972 base pair HindIII to BamHI restriction fragment of pSP64 (Promega).Fragment 1 is an approximately 1484 base pair Bgl II to AccI restrictionsub-fragment of the SPV HindIII restriction fragment M (23). Fragment 2is an approximately 1240 base pair HindIII fragment within the BamHI "d"fragment of EHV-1 (48). Fragment 3 is an approximately 3010 base pairBamHI to PvuII restriction fragment of plasmid pJF751 (11). Fragment 4is an approximately 2149 base pair AccI to HindIII subfragment of theSPV HindIII fragment M. The AccI sites in fragments 1 and 4 wereconverted to unique NotI sites using NotI linkers.

CLONING OF PARAINFLUENZA-3 VIRUS FUSION AND HEMAGGLUTININ GENES. Theparainfluenza-3 virus fusion (F) and hemagglutinin (HN) genes werecloned by a PCR CLONING procedure essentially as described by Katz etal. (42) for the HA gene of human influenza. Viral RNA prepared frombovine PI-3 virus grown in Madin-Darby bovine kidney (MDBK) cells wasfirst converted to cDNA utilizing an oligonucleotide primer specific forthe target gene. The cDNA was then used as a template for polymerasechain reaction (PCR) cloning (15) of the targeted region. The PCRprimers were designed to incorporate restriction sites which permit thecloning of the amplified coding regions into vectors containing theappropriate signals for expression in SPV. One pair of oligonucleotideswere required for each coding region. The F gene coding region from thePI-3 strain SF-4 (VR-281) was cloned using the following primers:5'-TTATGGATCCTGCTGCTGTGTTGAACAACTTTGT-3' (SEQ ID NO: 130) for cDNApriming and combined with 5'-CCGCGGATCCCATGACCATCACAACCATAATCATAGCC-3'(SEQ ID NO: 131) for PCR. The HN gene coding region from PI-3 strainSF-4 (VR-281) was cloned utilizing the following primers:5'-CGTCGGATCCCTTAGCTGCAGTTTTTTGGAACTTCTGTTTTGA-3' (SEQ ID NO: 132) forcDNA priming and combined with5'-CATAGGATCCCATGGAATATTGGAAACACACAAACAGCAC-3' (SEQ ID NO: 133) for PCR.Note that this general strategy is used to clone the coding region of Fand HN genes from other strains of PI-3. The DNA fragment for PI-3 HN orF was digested with BamHI to yield an 1730 bp or 1620 bp fragment,respectively. The PI-3 HN fragment is cloned into the BamHI site next tothe LP2EP2 promoter of the SPV homology vector. The PI-3 F fragment iscloned into the BamHI site next to the LP2EP2 promoter of the SPVhomology vector to yield homology vector 713-55.10.

CLONING OF BOVINE VIRAL DIARRHEA VIRUS gp48 and gp53 GENES. The bovineviral diarrhea gp48 and gp53 genes were cloned by a PCR CLONINGprocedure essentially as described by Katz et al. (42) for the HA geneof human influenza. Viral RNA prepared from BVD virus Singer straingrown in Madin-Darby bovine kidney (MDBK) cells was first converted tocDNA utilizing an oligonucleotide primer specific for the target gene.The cDNA was then used as a template for polymerase chain reaction (PCR)cloning (15) of the targeted region. The PCR primers were designed toincorporate restriction sites which permit the cloning of the amplifiedcoding regions into vectors containing the appropriate signals forexpression in SPV. One pair of oligonucleotides were required for eachcoding region. The gp48 gene coding region from the BVDV Singer strain(49) was cloned using the following primers:5'-ACGTCGGATCCCTTACCAAACCACGTCTTACTCTTGTTTTCC-3' (SEQ ID NO: 134) forcDNA priming and combined with5'-ACATAGGATCCCATGGGAGAAAACATAACACAGTGGAACC-3' (SEQ ID NO: 135) for PCR.The gp53 gene coding region from the BVDV Singer strain (49) was clonedusing the following primers: 5'-CGTGGATCCTCAATTACAAGAGGTATCGTCTAC-3'(SEQ ID NO: 136) for cDNA priming and combined with5'-CATAGATCTTGTGGTGCTGTCCGACTTCGCA-3' (SEQ ID NO: 137) for PCR. Notethat this general strategy is used to clone the coding region of gp48and gp53 genes from other strains of BVDV. The DNA fragment for BVDV gp48 was digested with BamHI to yield an 678 bp fragment. The DNA fragmentfor BVDV gp 53 was digested with BglII and BamHI to yield an 1187 bpfragment. The BVDV gp48 or gp53 DNA fragments were cloned into the BamHIsite next to the LP2EP2 promoter of the SPV homology vector to yieldhomology vectors, 727-78.1 and 738-96, respectively.

CLONING OF BOVINE RESPIRATORY SYNCYTIAL VIRUS FUSION, NUCLEOCAPSID ANDGLYCOPROTEIN GENES. The bovine respiratory syncytial virus fusion (F),nucleocapsid (N), and glycoprotein (G) genes were cloned by a PCRCLONING procedure essentially as described by Katz et al. (42) for theHA gene of human influenza. Viral RNA prepared from BRSV virus grown inbovine nasal turbinate (BT) cells was first converted to cDNA utilizingan oligonucleotide primer specific for the target gene. The cDNA wasthen used as a template for polymerase chain reaction (PCR) cloning (15)of the targeted region. The PCR primers were designed to incorporaterestriction sites which permit the cloning of the amplified codingregions into vectors containing the appropriate signals for expressionin SPV. One pair of oligonucleotides were required for each codingregion. The F gene coding region from the BRSV strain 375 (VR-1339) wascloned using the following primers:5'-TGCAGGATCCTCATTTACTAAAGGAAAGATTGTTGAT-3' (SEQ ID NO: 138) for cDNApriming and combined with 5'-CTCTGGATCCTACAGCCATGAGGATGATCATCAGC-3' (SEQID NO: 139) for PCR. The N gene coding region from BRSV strain 375(VR-1339) was cloned utilizing the following primers:5'-CGTCGGATCCCTCACAGTTCCACATCATTGTCTTTGGGAT-3' (SEQ ID NO: 140) for cDNApriming and combined with5'-CTTAGGATCCCATGGCTCTTAGCAAGGTCAAACTAAATGAC-3' (SEQ ID NO: 141) forPCR. The G gene coding region from BRSV strain 375 (VR-1339) was clonedutilizing the following primers:5'-CGTTGGATCCCTAGATCTGTGTAGTTGATTGATTTGTGTGA-3' (SEQ ID NO: 142) forcDNA priming and combined with5'-CTCTGGATCCTCATACCCATCATCTTAAATTCAAGACATTA-3' (SEQ ID NO: 143) forPCR. Note that this general strategy is used to clone the coding regionof F, N and G genes from other strains of BRSV. The DNA fragments forBRSV F, N, or G were digested with BamHI to yield 1722 bp, 1173 bp, or771 bp fragments, respectively. The BRSV F, N, and G DNA fragments werecloned into the BamHI site next to the LP2EP2 promoter of the SPVhomology vector to yield homology vectors, 727-20.10, 713-55.37 and727-20.5, respectively.

HOMOLOGY VECTOR 689-50.4 The plasmid 689-50.4 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliβ-galactosidase (lacz) marker gene and the infectious bursal diseasevirus (IBDV) polyprotein gene flanked by SPV DNA. Upstream of theforeign genes is an approximately 1484 base pair fragment of SPV DNA.Downstream of the foreign genes is ar. approximately 2149 base pairfragment of SPV DNA. When the plasmid is used according to theHOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV, avirus containing DNA coding for the foreign genes will result. Note thatthe β-galactosidase (lacZ) marker gene is under the control of asynthetic late pox promoter (LP1), and the IBDV polyprotein gene isunder the control of a synthetic late/early pox promoter (LP2EP2). Itmay be constructed utilizing standard recombinant DNA techniques (22,30), by joining restriction fragments from the following sources. Theplasmid vector is derived from an approximately 2972 base pair Hind IIIto BamHI restriction fragment of pSP64 (Promega). Fragment 1 is anapproximately 1484 base pair BglII to AccI restriction subfragment ofthe SPV HindIII restriction fragment M (23). Fragment 2 is anapproximately 3400 base pair fragment with SmaI and HpaI restrictionsites at the ends from plasmid 2-84/2-40 (51). This fragment containsthe IBDV polyprotein coding sequences. Fragment 3 is an approximately3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751(11). Fragment 4 is an approximately 2149 base pair AccI to HindIIIsubfragment of the SPV HindIII fragment M. The AccI sites in fragments 1and 4 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 689-50.7. The plasmid 689-50.7 was constructed for thepurpose of inserting foreign DNA into SPV. It incorporates an E. coliβ-galactosidase (lacZ) marker gene and the infectious bursal diseasevirus (IBDV) VP2 gene flanked by SPV DNA. Upstream of the foreign genesis an approximately 1484 base pair fragment of SPV DNA. Downstream ofthe foreign genes is an approximately 2149 base pair fragment of SPVDNA. When the plasmid is used according to the HOMOLOGOUS RECOMBINATIONPROCEDURE FOR GENERATING RECOMBINANT SPV, a virus containing DNA codingfor the foreign genes will result. Note that the β-galactosidase (lacZ)marker gene is under the control of a synthetic late pox promoter (LP1),and the IBDV VP2 gene is under the control of a synthetic late/early poxpromoter (LP2EP2). It may be constructed utilizing standard recombinantDNA techniques (22, 30), by joining restriction fragments from thefollowing sources. The plasmid vector is derived from an approximately2972 base pair HindIII to BamHI restriction fragment of pSP64 (Promega).Fragment 1 is an approximately 1484 base pair BglII to AccI restrictionsub-fragment of the SPV HindIII restriction fragment M (23). Fragment 2is an approximately 1081 base pair fragment with BclI and BamHIrestriction sites at the ends. This fragment codes for the IBDV VP2protein and is derived from a full length IBDV cDNA clone (51). Fragment3 is an approximately 3010 base pair BamHI to PvuII restriction fragmentof plasmid pJF751 (11). Fragment 4 is an approximately 2149 base pairAccI to HindIII sub-fragment of the SPV HindIII fragment M. The AccIsites in fragments 1 and 4 were converted to unique NotI sites usingNotI linkers.

EXAMPLES Example 1

Homology Vector 515-85.1. The homology vector 515-85.1 is a plasmiduseful for the insertion of foreign DNA into SPV. Plasmid 515-85.1contains a unique AccI restriction site into which foreign DNA may becloned. A plasmid containing such a foreign DNA insert may be usedaccording to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATINGRECOMBINANT SPV to generate a SPV containing the foreign DNA. For thisprocedure to be successful it is important that the insertion site(AccI) be in a region non-essential to the replication of the SPV andthat the site be flanked with swinepox virus DNA appropriate formediating homologous recombination between virus and plasmid DNAs. Wehave demonstrated that the AccI site in homology vector 515-85.1 may beused to insert foreign DNA into at least three recombinant SPV (seeexamples 2-4).

In order to define an appropriate insertion site, a library of SPVHindIII restriction fragments was generated. Several of theserestriction fragments (HindIII fragments G, J, and M see FIGS. 1A and1B) were subjected to restriction mapping analysis. Two restrictionsites were identified in each fragment as potential insertion sites.These sites included HpaI and NruI in fragment G, BalI and XbaI infragment J, and AccI and PstI in fragment M. A β-galactosidase (lacZ)marker gene was inserted in each of the potential sites. The resultingplasmids were utilized in the HOMOLOGOUS RECOMBINATION PROCEDURE FORGENERATING RECOMBINANT SPV. The generation of recombinant virus wasdetermined by the SCREEN FOR RECOMBINANT SPV EXPRESSING β-GALACTOSIDASEASSAYS. Four of the six sites were found to generate recombinant virus,however the ability of each of these viruses to be purified away fromthe parental SPV varied greatly. In one case virus could not be purifiedabove the level of 1%, in another case virus could not be purified abovethe level of 50%, and in a third case virus could not be purified abovethe level of 90%. The inability to purify these viruses indicatesinstability at the insertion site. This makes the corresponding sitesinappropriate for insertion of foreign DNA. However the insertion at onesite, the AccI site of Homology vector 515-85.1, resulted in a viruswhich was easily purified to 100% (see example 2), clearly defining anappropriate site for the insertion of foreign DNA.

The homology vector 515-85.1 was further characterized by DNA sequenceanalysis. Two regions of the homology vector were sequenced. The firstregion covers a 599 base pair sequence which flanks the unique AccI site(see FIGS. 2A and 2B). The second region covers the 899 base pairsupstream of the unique HindIII site (see FIGS. 2A and 2B). The sequenceof the first region codes for an open reading frame (ORF) which showshomology to amino acids 1 to 115 of the vaccinia virus (VV) O1L openreading frame identified by Goebel et al, 1990 (see FIGS. 3A, 3B and 3C). The sequence of the second region codes for an open reading framewhich shows homology to amino acids 568 to 666 of the same vacciniavirus O1L open reading frame (see FIGS. 3A, 3B and 3C). These datasuggest that the AccI site interrupts the presumptive VV O1L-like ORF atapproximately amino acid 41, suggesting that this ORF codes for a genenon-essential for SPV replication. Goebel et al. suggest that the VV O1LORF contains a leucine zipper motif characteristic of certain eukaryotictranscriptional regulatory proteins, however they indicate that it isnot known whether this gene is essential for virus replication. The DNAsequence located upstream of the VV 01L-like ORF (see FIG. 2A) would beexpected to contain a swinepox viral promoter. This swinepox viralpromoter will be useful as the control element of foreign DNA introducedinto the swinepox genome.

Example 2

S-SPV-003

S-SPV-003 is a swinepox virus that expresses a foreign gene. The genefor E.coli β-galactosidase (lacZ gene) was inserted into the SPV515-85.1 ORF. The foreign gene (lacz) is under the control of asynthetic early/late promoter (EP1LP2).

S-SPV-003 was derived from S-SPV-001 (Kasza strain). This wasaccomplished utilizing the homology vector 520-17.5 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-GALACTOSIDASE (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-003. This virus was assayed forβ-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable andexpressing the foreign gene. The assays described here were carried outin VERO cells as well as EMSK cells, indicating that VERO cells would bea suitable substrate for the production of SPV recombinant vaccines.S-SPV-003 has been deposited with the ATCC under Accession No. VR 2335.

Example 3

S-SPV-008

S-SPV-008 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacz gene) and the gene forpseudorabies virus (PRV) g50 (gpD) (26) were inserted into the SPV515-85.1 ORF. The lacZ gene is under the control of a synthetic latepromoter (LP1) and the g50 (gp)D gene is under the control of asynthetic early/late promoter (EP1LP2).

S-SPV-008 was derived from S-SPV-001 (Kasza strain). This wasaccomplished utilizing the homology vector 538-46.16 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-GALACTOSIDASE (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-008. This virus was assayed forβ-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable andexpressing the marker gene.

S-SPV-008 was assayed for expression of PRV specific antigens using theBLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV.Swine anti-PRV serum was shown to react specifically with S-SPV-008plaques and not with S-SPV-009 negative control plaques. All S-SPV-008observed plaques reacted with the swine antiserum indicating that thevirus was stably expressing the PRV foreign gene. The black plaque assaywas also performed on unfixed monolayers. The SPV plaques on the unfixedmonolayers also exhibited specific reactivity with swine anti-PRV serumindicating that the PRV antigen is expressed on the infected cellsurface.

To confirm the expression of the PRV g50 (gpD) gene product, cells wereinfected with SPV and samples of infected cell lysates were subjected toSDS-polyacrylamide gel electrophoresis. The gel was blotted and analyzedusing the WESTERN BLOTTING PROCEDURE. The swine anti-PRV serum was usedto detect expression of PRV specific proteins. As shown in FIG. 6, thelysate from S-SPV-008 infected cells exhibits a specific band ofapproximately 48 kd, the reported size of PRV g50 (gpD) (35).

PRV g50 (gpD) is the g50 (gpD) homologue of HSV-1 (26). Severalinvestigators have shown that VV expressing HSV-1 g50 (gpD) will protectmice against challenge with HSV-1 (6 and 34). Therefore the S-SPV-008should be valuable as a vaccine to protect swine against PRV disease.

It is anticipated that several other PRV glycoproteins will be useful inthe creation of recombinant swinepox vaccines to protect against PRVdisease. These PRV glycoproteins include gpII (28), gpIII (27), and gpH(19). The PRV gpIII coding region has been engineered behind severalsynthetic pox promoters. The techniques utilized for the creation ofS-SPV-008 will be used to create recombinant swinepox viruses expressingall four of these PRV glycoprotein genes. Such recombinant swinepoxviruses will be useful as vaccines against PRV disease. Since the PRVvaccines described here do not express PRV gpX or gpI, they would becompatible with current PRV diagnostic tests (gX HerdChek®, gI HerdChek®and ClinEase®) which are utilized to distinguish vaccinated animals frominfected animals. S-SPV-008 has been deposited with the ATCC underAccession No. VR 2339.

Example 4

S-SPV- 011

S-SPV-011 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli B-galactosidase (lacZ) and the gene forpseudorabies virus gIII (gpC) were inserted into the unique PstIrestriction site (PstI linkers inserted into a unique AccI site) of thehomology vector 570-33.32. The lac Z gene is under the control of thesynthetic late promoter (LP1) and the PRV gIII (gpC) gene is under thecontrol of the synthetic early promoter (EP2).

S-SPV-011 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 570-91.21 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING B-GALACTOSIDASE (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-011. This virus was assayed forB-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

S-SPV-011 was assayed for expression of PRV specific antigens using theBLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV.Polyclonal goat anti-PRV gIII (gpC) antibody was shown to reactspecifically with S-SPV-011 plaques and not with S-SPV-001 negativecontrol plaques. All S-SPV-011 observed plaques reacted with the swineanti-PRV serum indicating that the virus was stably expressing the PRVforeign gene. The assays described here were carried out in EMSK cells,indicating that EMSK cells would be a suitable substrate for theproduction of SPV recombinant vaccines.

To confirm the expression of the PRV gIII (gpC) gene product, cells wereinfected with SPV and samples of infected cell lysates were subjected toSDS-polyacrylamide gel electrophoresis. The gel was blotted and analyzedusing the WESTERN BLOTTING PROCEDURE. Polyclonal goat anti-PRV gIII(gpC) antibody was used to detect expression of PRV specific proteins.As shown in FIG. 16, the lysate from S-SPV-011 infected cells exhibits aspecific band of approximately 92 kd, the reported size of PRV gIII(gpC) (37).

Recombinant-expressed PRV gIII (gpC) has been shown to elicit asignificant immune response in mice and swine (37, 38). Furthermore,when gIII (gpC) is coexpressed with gII (gpB) or g50 (gpD), significantprotection from challenge with virulent PRV is obtained (39). ThereforeS-SPV-011 should be valuable as a vaccine to protect swine against PRVdisease. Since the PRV vaccines described here do not express PRV gpX orgpI, they would be compatible with current PRV diagnostic tests (gXHerdChek®, gI HerdChek® and ClinEase®) which are utilized to distinguishvaccinated animals from infected animals.

Example 5

S-SPV-012

S-SPV-012 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli B-galactosidase (lacz) and the gene forpseudorabies virus gIII (gpC) were inserted into the unique PstIrestriction site (PstI linkers inserted into a unique AccI site) of thehomology vector 570-33.32. The lacZ gene is under the control of thesynthetic late promoter (LP1) and the PRV gIII (gpC) gene is under thecontrol of the synthetic early late promoter (EP1LP2).

S-SPV-012 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 570-91.41 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING B-GALACTOSIDASE (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-012. This virus was assayed forB-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

S-SPV-012 was assayed for expression of PRV specific antigens using theBLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV.Polyclonal goat anti-PRV gIII (gpC) antibody was shown to reactspecifically with S-SPV-012 plaques and not with S-SPV-001 negativecontrol plaques. All S-SPV-012 observed plaques reacted with the swineanti-PRV serum, indicating that the virus was stably expressing the PRVforeign gene. The assays described here were carried out in EMSK andVERO cells, indicating that EMSK cells would be a suitable substrate forthe production of SPV recombinant vaccines.

To confirm the expression of the PRV gIII (gpC) gene product, cells wereinfected with S-SPV-012 and samples of infected cell lysates weresubjected to SDS-polyacrylamide gel electrophoresis. The gel was blottedand analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal goatanti-PRV gIII (gpC) antibody was used to detect expression of PRVspecific proteins. As shown in FIG. 16, the lysate from S-SPV-012infected cells exhibits two specific bands which are the reported sizeof PRV gIII (gpC) (37)--a 92 kd mature form and a 74 kd pre-golgi form.

Recombinant-expressed PRV gIII (gpC) has been shown to elicit asignificant immune response in mice and swine (37, 38). Furthermore,when gIII (gpC) is coexpressed with gII (gpB) or g50 (gpD), significantprotection from challenge with virulent PRV is obtained (39). ThereforeS-SPV-012 should be valuable as a vaccine to protect swine against PRVdisease. Since the PRV vaccines described here do not express PRV gpX orgpI, they would be compatible with current PRV diagnostic tests (gXHerdChek®, gI HerdChek® and ClinEase®) which are utilized to distinguishvaccinated animals from infected animals.

Example 6

S-SPV-013

S-SPV-013 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli B-galactosidase (lacZ) and the gene forpseudorabies virus gill (gpC) were inserted into the unique PstIrestriction site (PstI linkers inserted into a unique AccI site) of thehomology vector 570-33.32. The lacZ gene is under the control of thesynthetic late promoter (LP1) and the PRV gIII (gpC) gene is under thecontrol of the synthetic late early promoter (LP2EP2).

S-SPV-013 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 570-91.64 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING B-GALACTOSIDASE (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-013. This virus was assayed forB-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

S-SPV-013 was assayed for expression of PRV specific antigens using theBLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV.Polyclonal goat anti-PRV gIII (gpC) antibody was shown to reactspecifically with S-SPV-013 plaques and not with S-SPV-001 negativecontrol plaques. All S-SPV-013 observed plaques reacted with the swineanti-PRV serum indicating that the virus was stably expressing the PRVforeign gene. The assays described here were carried out in EMSK andVERO cells, indicating that EMSK cells would be a suitable substrate forthe production of SPV recombinant vaccines.

To confirm the expression of the PRV gIII (gpC) gene product, cells wereinfected with SPV and samples of infected cell lysates were subjected toSDS-polyacrylamide gel electrophoresis. The gel was blotted and analyzedusing the WESTERN BLOTTING PROCEDURE. Polyclonal goat anti-PRV gIII(gpC) antibody was used to detect expression of PRV specific proteins.As shown in FIG. 16, the lysate from S-SPV-013 infected cells exhibitstwo specific bands which are the reported size of PRV gIII (gpC) (37)--a92 kd mature form and a 74 kd pre-Golgi form.

Recombinant-expressed PRV gIII (gpC) has been shown to elicit asignificant immune response in mice and swine (37, 38). Furthermore,when gIII (gpC) is coexpressed with gII (gpB) or g50 (gpD), significantprotection from challenge with virulent PRV is obtained. (39) ThereforeS-SPV-013 should be valuable as a vaccine to protect swine against PRVdisease. Since the PRV vaccines described here do not express PRV gpX orgpI, they would be compatible with current PRV diagnostic tests (gXHerdChek®, gI HerdChek® and ClinEase®) which are utilized to distinguishvaccinated animals from infected animals. S-SPV-013 has been depositedwith the ATCC.

Protection against Aujeszky's disease using recombinant swinepox virusvaccines S-SPV-008 and S-SPV-013.

A vaccine containing S-SPV-008 and S-SPV-013 (1 x 106PFU/ml) (2 ml of a1:1 mixture of the two viruses) was given to two groups of pigs (5 pigsper group) by intradermal inoculation or by oral/pharyngeal spray. Acontrol group of 5 pigs received S-SPV-001 by both intradermal andoral/pharyngeal inoculation. Pigs were challenged three weekspost-vaccination with virulent PRV, strain 4892, by intranasalinoculation. The table presents a summary of clinical responses. Thedata support an increase in protection against Aujeszky's disease in theS-SPV-008/S-SPV-013 vaccinates compared to the S-SPV-001 vaccinatecontrols.

    __________________________________________________________________________                 Post-challenge                                                                              Post-challenge                                         Respiratory Post-challenge Group average:                                     Signs: CNS signs: (Days of                                                   Route of (# with signs/ (# with signs/ clinical                              Vaccine inoculation total number) total number) signs)                      __________________________________________________________________________    S-SPV-008 +                                                                         Intradermal                                                                          3/5    0/5    2.6                                                  S-SPV-013                                                                     S-SPV-008 + Oral/ 3/5 0/5 2.2                                                 S-SPV-013 pharyngeal                                                          S-SPV-001 Intradermal + 5/5 2/5 7.8                                           (Control) Oral/                                                                Pharyngeal                                                                 __________________________________________________________________________

Example 7

S-SPV-015

S-SPV-015 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene forpseudorabies virus (PRV) gII (gpB) were inserted into the SPV 617-48.1ORF (a unique NotI restriction site has replaced a unique AccI restriction site). The lacZ gene is under the control of the synthetic latepromoter (LP1), and the PRV gB gene is under the control of thesynthetic late/early promoter (LP2EP2).

S-SPV-015 was derived from S-SPV-001 (Kasza Strain) . This wasaccomplished utilizing the homology vector 727-54.60 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-015. This virus was assayed forβ-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

S-SPV-015 was assayed for expression of PRV-specific antigens using theBLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV.Polyclonal swine anti-PRV serum was shown to react specifically withS-SPV-015 plaques and not with S-SPV-001 negative control plaques. AllS-SPV-015 observed plaques reacted with the antiserum indicating thatthe virus was stably expressing the PRV foreign gene. The assaysdescribed here were carried out in ESK-4 cells, indicating that ESK-4cells would be a suitable substrate for the production of SPVrecombinant vaccines.

To confirm the expression of the PRV gII gene product, cells wereinfected with SPV-015 and samples of infected cell lysates weresubjected to SDS-polyacrylamide gel electrophoresis. The gel was blottedand analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal swineanti-PRV serum was used to detect expression of PRV specific proteins.The lysate from S-SPV-015 infected cells exhibited bands correspondingto 120 kd, 67 kd and 58 kd, which are the expected size of the PRV gIIglycoprotein.

S-SPV-015 is useful as a vaccine in swine against pseudorabies virus. Asuperior vaccine is formulated by combining S-SPV-008 (PRV g50),S-SPV-013 (PRV gIII), and S-SPV-015 for protection against pseudorabiesin swine.

Therefore S-SPV-015 should be valuable as a vaccine to protect swineagainst PRV disease. Since the PRV vaccines described here do notexpress PRV gpX or gpI, they would be compatible with current PRVdiagnostic tests (gX HerdChek®, gI HerdChek® and ClinEase®) which areutilized to distinguish vaccinated animals from infected animals.S-SPV-015 has been deposited with the ATCC.

Example 8

Recombinant swinepox virus expressing more than one pseudorabies virus(PRV) glycoproteins, which can elicit production of neutralizingantibodies against pseudorabies virus, is constructed in order to obtaina recombinant swinepox virus with enhanced ability to protect againstPRV infection than that which can be obtained by using a recombinantswinepox virus expressing only one of those PRV glycoproteins.

There are several examples of such recombinant swinepox virus expressingmore than one PRV glycoproteins: a recombinant swinepox virus expressingPRV g50 (gpD) and gIII (gpC), a recombinant swinepox virus expressingPRV g50 (gpD) and gII (gpB); a recombinant swinepox virus expressing PRVgII (gpB) and gIII (gpC); and a recombinant swinepox virus expressingPRV g50 (gpD), gIII (gpC) and gII (gpB). Each of the viruses cited aboveis also engineered to contain and express E. coli B-galactosidase (lacZ) gene, which will facilitate the cloning of the recombinant swinepoxvirus.

Listed below are three examples of a recombinant swinepox virusexpressing PRV g50 (gpD), PRV gIII (gpC), PRV gII (gpB) and E. coliB-galactosidase (lacZ):

a) Recombinant swinepox virus containing and expressing PRV g50 (gpD)gene, PRV gIII (gpC) gene, PRV gII (gpB) gene and lacZ gene. All fourgenes are inserted into the unique AccI restriction endonuclease sitewithin the HindIII M fragment of the swinepox virus genome. PRV g50(gpD) gene is under the control of a synthetic early/late promoter(EP1LP2), PRV gIII (gpC) gene is under the control of a synthetic earlypromoter (EP2), PRV gII (gpB) gene is under the control of a syntheticlate/early promoter (LP2EP2) and lacZ gene is under the control of asynthetic late promoter (LP1).

b) Recombinant swinepox virus containing and expressing PRV g50 (gpD)gene, PRV gIII (gpC) gene, PRV gII (gpB) gene and lacz gene. All fourgenes are inserted into the unique AccI restriction endonuclease sitewithin the HindIII M fragment of the swinepox virus genome. PRV g50(gpD) gene is under the control of a synthetic early/late promoter(EP1LP2), PRV gIII (gpC) gene is under the control of a syntheticearly/late promoter (EP1LP2), PRV gII (gpB) gene is under the control ofa synthetic late/early promoter (LP2EP2) and lacZ gene is under thecontrol of a synthetic late promoter (LP1).

c) Recombinant swinepox virus containing and expressing PRV g50 (gpD)gene, PRV gIII (gpC) gene, PRV gII (gpB) gene and lacZ gene. All fourgenes are inserted into the unique AccI restriction endonuclease sitewithin the HindIII M fragment of the swinepox virus genome. PRV g50(gpD) gene is under the control of a synthetic early/late promoter(EP1LP2), PRV gIII (gpC) gene is under the control of a syntheticlate/early promoter (LP2EP2), PRV gII (gpB) gene is under the control ofa synthetic late/early promoter (LP2EP2) and lacZ gene is under thecontrol of a synthetic late promoter (LP1).

Example 9

S-SPV-009

S-SPV-009 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ gene) and the gene forNewcastle's Disease virus hemagglutinin (HN) gene were inserted into theSPV 515-85.1 ORF. The lacZ gene is under the control of a synthetic latepromoter (LP1) and the HN gene is under the control of an syntheticearly/late promoter (EP1LP2).

S-SPV-009 was derived from S-SPV-001 (Kasza strain). This wasaccomplished utilizing the homology vector 538-46.26 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-GALACTOSIDASE (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-009. This virus was assayed forβ-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable andexpressing the marker gene.

S-SPV-009 was assayed for expression of PRV specific antigens using theBLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV.Rabbit anti-NDV HN serum was shown to react specifically with S-SPV-009plaques and not with S-SPV-008 negative control plaques. All S-SPV-009observed plaques reacted with the swine antiserum indicating that thevirus was stably expressing the NDV foreign gene. S-SPV-009 has beendeposited with the ATCC under Accession No. VR 2344).

To confirm the expression of the NDV HN gene product, cells wereinfected with SPV and samples of infected cell lysates were subjected toSDS-polyacrylamide gel electrophoresis. The gel was blotted and analyzedusing the WESTERN BLOTTING PROCEDURE. The rabbit anti-NDV HN serum wasused to detect expression of the HN protein. The lysate from S-SPV-009infected cells exhibited a specific band of approximately 74 kd, thereported size of NDV HN (29).

Example 10

S-SPV-014

S-SPV-014 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli B-galactosidase (lacZ) and the gene for infectiouslaryngotracheitis virus glycoprotein G (ILT gpG) were inserted into theSPV 570-33.32 ORF (a unique PstI site has replaced the unique AccIsite). The lacZ gene is under the control of the synthetic late promoter(LP1), and the ILT gpG gene is under the control of the syntheticearly/late promoter (EP1LP2).

S-SPV-014 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 599-65.25 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING B-GALACTOSIDASE (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-014. This virus was assayed forB-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene. The assays described here were carried outin ESK-4 cells, indicating that ESK-4 cells would be a suitablesubstrate for the production of SPV recombinant vaccines.

To confirm the expression of the ILT gpG gene product, cells wereinfected with SPV-014 and samples of infected cell lysates weresubjected to SDS-polyacrylamide gel electrophoresis. The gel was blottedand analyzed using the WESTERN BLOTTING PROCEDURE. Peptide antisera toILT gG was used to detect expression of ILT specific proteins. Thelysate from S-SPV-014 infected cells exhibited a band at 43 kd which isthe expected size of the ILT gpG protein and additional bands of highermolecular weight which represent glycosylated forms of the protein whichare absent in deletion mutants for ILT gpG.

This virus is used as an expression vector for expressing ILTglycoprotein G (gpG). Such ILT gpG is used as an antigen to identifyantibodies directed against the wild-type ILT virus as opposed toantibodies directed against gpG deleted ILT viruses. This virus is alsoused as an antigen for the production of ILT gpG specific monoclonalantibodies. Such antibodies are useful in the development of diagnostictests specific for the ILT gpG protein. Monoclonal antibodies aregenerated in mice utilizing this virus according to the PROCEDURE FORPURIFICATION OF VIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS (Materials &Methods).

Example 11

S-SPV-016

S-SPV-016 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli B-galactosidase (lacZ) and the gene for infectiouslaryngotracheitis virus glycoproteinI (ILT gpI) were inserted into theSPV 617-48.1 ORF (a unique NotI restriction site has replaced a uniqueAccI restriction site). The lacZ gene is under the control of thesynthetic late promoter (LP1), and the ILT gpI gene is under the controlof the synthetic late/early promoter (LP2EP2).

S-SPV-016 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 624-20.1C (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING B-GALACTOSIDASE (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-016. This virus was assayed forB-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, al; plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

S-SPV-016 was assayed for expression of ILT gpI- andB-galactosidase-specific antigens using the BLACK PLAQUE SCREEN FORFOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Polyclonal chicken anti-ILTantibody was shown to react specifically with S-SPV-016 plaques and notwith S-SPV-017 negative control plaques. All S-SPV-016 observed plaquesreacted with the chicken antiserum indicating that the virus was stablyexpressing the ILT foreign gene. The assays described here were carriedout in ESK-4 cells, indicating that ESK-4 cells would be a suitablesubstrate for the production of SPV recombinant vaccines.

To confirm the expression of the ILT gpI gene product, cells wereinfected with SPV-016 and samples of infected cell lysates weresubjected to SDS-polyacrylamide gel electrophoresis. The gel was blottedand analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal chickenanti-ILT antibody was used to detect expression of ILT specificproteins. The lysate from S-SPV-016 infected cells exhibits a range ofbands reactive to the anti-ILT antibody from 40 to 200 kd indicatingthat the ILT gpI may be heavily modified.

This virus is used as an expression vector for expressing ILTglycoprotein I (gpI). Such ILT gpI is used as an antigen to identifyantibodies directed against the wild-type ILT virus as opposed toantibodies directed against gpI deleted ILT viruses. This virus is alsoused as an antigen for the production of ILT gpI specific monoclonalantibodies. Such antibodies are useful in the development of diagnostictests specific for the ILT gpI protein. Monoclonal antibodies aregenerated in mice utilizing this virus according to the PROCEDURE FORPURIFICATION OF VIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS (Materials &Methods)

Example 12

S-SPV-017

S-SPV-017 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli B-galactosidase (lacZ) and the gene for infectiousbovine rhinotracheitis virus glycoprotein G (IBR gpG) were inserted intothe SPV 617-48.1 ORF (a unique NotI restriction site has replaced aunique AccI restriction site). The lacz gene is under the control of thesynthetic late promoter (LP1), and the IBR gpG gene is under the controlof the synthetic late/early promoter (LP2EP2).

S-SPV-017 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 614-83.18 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING B-GALACTOSIDASE (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-017. This virus was assayed forB-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

S-SPV-017 was assayed for expression of IBR-specific antigens using theBLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV.Monoclonal antibodies and peptide antisera to IBR gpG were shown toreact specifically with S-SPV-017 plaques and not with S-SPV-016negative control plaques. All S-SPV-017 observed plaques reacted withthe antiserum indicating that the virus was stably expressing the IBRforeign gene. The assays described here were carried out in ESK-4 cells,indicating that ESK-4 cells would be a suitable substrate for theproduction of SPV recombinant vaccines.

To confirm the expression of the IBR gpG gene product, cells wereinfected with SPV-017 and samples of infected cell lysates weresubjected to SDS-polyacrylamide gel electrophoresis. The gel was blottedand analyzed using the WESTERN BLOTTING PROCEDURE. Antisera to IBR gpGwas used to detect expression of IBR specific proteins. The lysate fromS-SPV-017 infected cells exhibited a band at 43 kd which is the expectedsize of the IBR gpG protein and additional bands of higher molecularweight which represent glycosylated forms of the protein which areabsent in deletion mutants for IBR gpG.

This virus is used as an expression vector for expressing IBRglycoprotein G (gpG). Such IBR gpG is used as an antigen to identifyantibodies directed against the wild-type IBR virus as opposed toantibodies directed against gpG deleted IBR viruses. This virus is alsoused as an antigen for the production of IBR gpG specific monoclonalantibodies. Such antibodies are useful in the development of diagnostictests specific for the IBR gpG protein. Monoclonal antibodies aregenerated in mice utilizing this virus according to the PROCEDURE FORPURIFICATION OF VIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS (Materials &Methods).

Example 13

S-SPV-019

S-SPV-019 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene for infectiousbovine rhinotracheitis virus (IBRV) gE were inserted into the SPV617-48.1 ORF (a unique NotI restriction site has replaced a unique AccIrestriction site). The lacZ gene is under the control of the syntheticlate promoter (LP1), and the IBRV gE gene is under the control of thesynthetic late/early promoter (LP2EP2).

S-SPV-019 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 708-78.9 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-019. This virus was assayed forβ-galactosidase expression, purity and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

This virus is used as an expression vector for expressing IBRglycoprotein E (gpE). Such IBR gpE is used as an antigen to identifyantibodies directed against the wild-type IBR virus as opposed toantibodies directed against gpE deleted IBR viruses. This virus is alsoused as an antigen for the production of IBR gpE specific monoclonalantibodies. Such antibodies are useful in the development of diagnostictests specific for the IBR gpE protein. Monoclonal antibodies aregenerated in mice utilizing this virus according to the PROCEDURE FORPURIFICATION OF VIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS (Materials &Methods).

Example 14

S-SPV-018

S-SPV-018 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli B-galactosidase (lacZ) and the gene forpseudorabies virus glycoprotein E (PRV gpE) are inserted into the SPV570-33.32 ORF (a unique PstI site has replaced the unique AccI site).The lacZ gene is under the control of the synthetic late promoter (LP1),and the PRV gpE gene is under the control of the synthetic early/latepromoter (EP1LP2).

S-SPV-018 is derived from the S-SPV-001 (Kasza Strain). This isaccomplished utilizing the final homology vector and virus S-SPV-001 inthe HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV.The transfection stock is screened by the SCREEN FOR RECOMBINANT SPVEXPRESSING B-GALACTOSIDASE (BLUOGAL AND CPRG ASSAYS). Red plaquepurification of the recombinant virus is designated S-SPV-018. Thisvirus is assayed for B-galactosidase expression, purity, and insertstability by multiple passages monitored by the blue plaque assaydescribed in Materials and Methods. After the initial three rounds ofpurification, all plaques observed are blue indicating that the virus ispure, stable, and expressing the foreign gene.

This virus is used as an expression vector for expressing PRVglycoprotein E (gpE). Such PRV gpE is used as an antigen to identifyantibodies directed against the wild-type PRV virus as opposed toantibodies directed against gpE deleted PRV viruses. This virus is alsoused as an antigen for the production of PRV gpE specific monoclonalantibodies. Such antibodies are useful in the development of diagnostictests specific for the PRV gpE protein. Monoclonal antibodies aregenerated in mice utilizing this virus according to the PROCEDURE FORPURIFICATION OF VIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS (Materials &Methods).

Example 15

Homology Vector 520-90.15

The homology vector 520-90.15 is a plasmid useful for the insertion offoreign DNA into SPV. Plasmid 520-90.15 contains a unique NdeIrestriction site into which foreign DNA may be cloned. A plasmidcontaining such a foreign DNA insert has been used according to theHOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV togenerate a SPV containing the foreign DNA. For this procedure to besuccessful, it is important that the insertion site be in a regionnon-essential to the replication of the SPV and that the site be flankedwith swinepox virus DNA appropriate for mediating homologousrecombination between virus and plasmid DNAs. The unique NdeIrestriction site in plasmid 520-90.15 is located within the codingregion of the SPV thymidine kinase gene (32). Therefore, we have shownthat the thymidine kinase gene of swinepox virus is non-essential forDNA replication and is an appropriate insertion site.

Example 16

S-PRV-010

S-SPV-010 is a swinepox virus that expresses a foreign gene. The E. coliB-galactosidase (lacz) gene is inserted into a unique NdeI restrictionsite within the thymidine kinase gene. The foreign gene (lacZ) is underthe control of the synthetic late promoter, LP1. We have shown that theswinepox virus thymidine kinase gene is non-essential for replication ofthe virus and is an appropriate insertion site.

A 1739 base pair HindIII-BamHI fragment subcloned from the HindIII Gfragment contains the swinepox virus thymidine kinase gene and isdesignated homology vector 520-90.15. The homology vector 520-90.15 wasdigested with Nde I, and AscI linkers were inserted at this unique sitewithin the thymidine kinase gene. The LP1 promoter-lac Z cassette withAscI linkers was ligated into the Asc I site within the thymidine kinasegene. The recombinant homology vector 561-36.26 was cotransfected withvirus S-SPV-001 by the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATINGRECOMBINANT SPV and virus plaques expressing B-galactosidase wereselected by SCREEN FOR RECOMBINANT SPV EXPRESSING B-GALACTOSIDASE(BLUOGAL AND CPRG ASSAY). The final result of blue and red plaquepurification was the recombinant virus designated S-SPV-010. This viruswas assayed for B-galactosidase expression, purity, and insert stabilityby multiple passages monitored by the blue plaque assay as described inMaterials and Methods. After the initial three rounds of purification,all plaques observed were blue indicating that the virus was pure,stable and expressing the foreign gene. The assays described here werecarried out in ESK-4 cells, indicating that ESK-4 cells would be asuitable substrate for the production of SPV recombinant vaccines.

Example 17

The development of vaccines utilizing the swinepox virus to expressantigens from various disease causing microorganisms can be engineered.

Transmissible Gastroenteritis Virus

The major neutralizing antigen of the transmissible gastroenteritisvirus (TGE), glycoprotein 195, for use in the swinepox virus vector hasbeen cloned. The clone of the neutralizing antigen is disclosed in U.S.Ser. No. 078,519, filed Jul. 27, 1987. It is contemplated that theprocedures that have been used to express PRV g50 (gpD) in SPV and aredisclosed herein are applicable to TGE.

Porcine Parvovirus

We have cloned the major capsid protein of the porcine (swine)parvovirus (PPV) for use in the swinepox virus vector. The clone of thecapsid protein is disclosed in U.S. Pat. No. 5,068,192 issued Nov. 26,1991. It is contemplated that the procedures that have been used toexpress PRV g50 (gpD) in SPV and are disclosed herein are applicable toPPV.

Swine Rotavirus

We have cloned the major neutralizing antigen of the swine rotavirus,glycoprotein 38, for use in the swinepox virus vector. The clone ofglycoprotein 38 is disclosed in U.S. Pat. No. 5,068,192 issued Nov. 26,1991. It is contemplated that the procedures that have been used toexpress PRV g50 (gpD) in SPV and are disclosed herein are applicable toSRV.

Hog Cholera Virus

The major neutralizing antigen of the bovine viral diarrhea (BVD) viruswas cloned as disclosed in U.S. Ser. No. 225,032, filed Jul. 27, 1988.Since the BVD and hog cholera viruses are cross protective (31), the BVDvirus antigen has been targeted for use in the swinepox virus vector. Itis contemplated that the procedures that have been used to express PRVg50 (gpD) in SPV and are disclosed herein are applicable to BVD virus.

Serpulina hyodysenteriae

A protective antigen of Serpulina hyodysenteriae (3), for use in theswinepox virus vector has been cloned. It is contemplated that theprocedures that have been used to express PRV gp50 in SPV and aredisclosed herein are also applicable to Serpulina hyodysenteriae.

Antigens from the following microorganisms may also be utilized todevelop animal vaccines: swine influenza virus, foot and mouth diseasevirus, African swine fever virus, hog cholera virus, Mycoplasmahyopneumoniae, porcine reproductive and respiratory syndrome/swineinfertility and respiratory syndrome (PRRS/SIRS).

Antigens from the following microorganisms may also be utilized todevelop animal vaccines: feline leukemia virus, feline immunodeficiencyvirus, feline herpesvirus, feline infectious peritonitis virus, canineherpesvirus, canine coronavirus, canine parvovirus, parasitic diseasesin animals (including Dirofilaria immitis in dogs and cats), equineinfectious anemia, Streptococcus equi, coccidia, emeria, chicken anemiavirus, Borrelia bergdorferi, bovine coronavirus, pasteurella,haemolytica.

Example 18

Recombinant swinepox viruses express equine influenza virus typeA/Alaska 91, equine influenza virus type A/Prague 56, equine herpesvirustype 1 gB, or equine herpesvirus type 1 gD genes. S-SPV-033 andS-SPV-034 are useful as vaccines against equine influenza infection, andS-SPV-038 and S-SPV-039 are useful as a vaccine against equineherpesvirus infection which causes equine rhinotracheitis and equineabortion. These equine influenza and equine herpesvirus antigens are keyto raising a protective immune response in the animal. The recombinantviruses are useful alone or in combination as an effective vaccine. Theswinepox virus is useful for cloning other subtypes of equine influenzavirus (including EIVA/Miami/63 and EIVA/Kentucky/81) to protect againstrapidly evolving variants in this disease. S-SPV-033, S-SPV-034,S-SPV-038, and S-SPV-039 are also useful as an expression vector forexpressing equine influenza or equine herpesvirus antigens. Such equineinfluenza or equine herpesvirus antigens are useful to identifyantibodies directed against the wild-type equine influenza virus orequine herpesvirus. The viruses are also useful to in producing antigensfor the production of monospecific polyclonal or monoclonal antibodies.Such antibodies are useful in the development of diagnostic testsspecific for the viral proteins. Monoclonal or polyclonal antibodies aregenerated in mice utilizing these viruses according to the PROCEDURE FORPURIFICATION OF VIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS (Materialsand Methods).

Example 18A

S-SPV-033:

S-SPV-033 is a recombinant swinepox virus that expresses at least twoforeign genes. The gene for E. coli β-galactosidase (lacZ) and the genefor equine influenza virus type A/Alaska 91 neuraminidase were insertedinto the SPV 617-48.1 ORF (a unique NotI restriction site has replaced aunique AccI restriction site). The lacZ gene is under the control of thesynthetic late promoter (LP1), and the EIV AK/91 NA gene is under thecontrol of the synthetic late/early promoter (LP2EP2).

S-SPV-033 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 732-18.4 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-033. This virus was assayed forβ-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

Example 18B

S-SPV-034:

S-SPV-034 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene for equineinfluenza virus type A/Prague 56 neuraminidase were inserted into theSPV 617-48.1 ORF (a unique NotI restriction site has replaced a uniqueAccI restriction site). The lacZ gene is under the control of thesynthetic late promoter (LP1), and the EIV PR/56 NA gene is under thecontrol of the synthetic late/early promoter (LP2EP2).

S-SPV-034 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 723-59A9.22 (see Materialsand Methods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATIONPROCEDURE FOR GENERATING RECOMBINANT SPV. The transfection stock wasscreened by the SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase(BLUOGAL AND CPRG ASSAYS). The final result of red plaque purificationwas the recombinant virus designated S-SPV-034. This virus was assayedfor β-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

S-SPV-034 was assayed for expression of EIV-specific antigens using theBLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV.Monospecific polyclonal antibodies to EIV PR/56 NA were shown to reactspecifically with S-SPV-034 plaques and not with S-SPV-001 negativecontrol plaques. All S-SPV-034 observed plaques reacted with theantiserum indicating that the virus was stably expressing the EIV PR/56NA gene. The assays described here were carried out in ESK-4 cells,indicating that ESK-4 cells would be a suitable substrate for theproduction of SPV recombinant vaccines.

Example 18C

S-SPV-038:

S-SPV-038 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene for equineherpesvirus type 1 glycoprotein B are inserted into the SPV 617-48.1 ORF(a unique NotI restriction site has replaced a unique AccI restrictionsite). The lacZ gene is under the control of the synthetic late promoter(LP1), and the EHV-1 gB gene is under the control of the syntheticlate/early promoter (LP2EP2).

S-SPV-038 is derived from S-SPV-001 (Kasza Strain). This is accomplishedutilizing the homology vector 744-34 (see Materials and Methods) andvirus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATINGRECOMBINANT SPV. The transfection stock is screened by the SCREEN FORRECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL AND CPRG ASSAYS).The final result of red plaque purification is the recombinant virusdesignated S-SPV-038. This virus is assayed for β-galactosidaseexpression, purity, and insert stability by multiple passages monitoredby the blue plaque assay as described in Materials and Methods. Afterthe initial three rounds of purification, all plaques observed are blueindicating that the virus is pure, stable, and expressing the foreigngene.

Example 18D

S-SPV-039:

S-SPV-039 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene for equineherpesvirus type 1 glycoprotein D are inserted into the SPV 617-48.1 ORF(a unique NotI restriction site has replaced a unique AccI restrictionsite). The lacZ gene is under the control of the synthetic late promoter(LP1), and the EHV-1 gD gene is under the control of the syntheticlate/early promoter (LP2EP2).

S-SPV-039 is derived from S-SPV-001 (Kasza Strain). This is accomplishedutilizing the homology vector 744-38 (see Materials and Methods) andvirus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATINGRECOMBINANT SPV. The transfection stock is screened by the SCREEN FORRECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL AND CPRG ASSAYS).The final result of red plaque purification is the recombinant virusdesignated S-SPV-039. This virus is assayed for β-galactosidaseexpression, purity, and insert stability by multiple passages monitoredby the blue plaque assay as described in Materials and Methods. Afterthe initial three rounds of purification, all plaques observed are blueindicating that the virus is pure, stable, and expressing the foreigngene.

Example 19

Recombinant swinepox viruses express bovine respiratory syncytial virusattachment protein (BRSV G), BRSV Fusion protein (BRSV F), BRSVnucleocapsid protein (BRSV N), bovine viral diarrhea virus (BVDV) gp48,BVDV gp53, bovine parainfluenza virus type 3 (BPI-3) F, or BPI-3 HN.S-SPV-020, S-SPV-029, S-SPV-030, and S-SPV-032, S-SPV-028 are useful asvaccines against bovine disease. These BRSV, BVDV, and BPI-3 antigensare key to raising a protective immune response in the animal. Therecombinant viruses are useful alone or in combination as an effectivevaccine. The swinepox virus is useful for cloning other subtypes ofBRSV, BVDV, and BPI-3 to protect against rapidly evolving variants inthis disease. S-SPV-020, S-SPV-029, S-SPV-030, and S-SPV-032, S-SPV-028are also useful as an expression vector for expressing BRSV, BVDV, andBPI-3 antigens. Such BRSV, BVDV, and BPI-3 antigens are useful toidentify antibodies directed against the wild-type BRSV, BVDV, andBPI-3. The viruses are also useful as antigens for the production ofmonospecific polyclonal or monoclonal antibodies. Such antibodies areuseful in the development of diagnostic tests specific for the viralproteins. Monoclonal or polyclonal antibodies are generated in miceutilizing these viruses according to the PROCEDURE FOR PURIFICATION OFVIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS (Materials and Methods).

Example 19A

S-SPV-020:

S-SPV-020 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene for bovinerespiratory syncytial virus (BRSV) G were inserted into the SPV 617-48.1ORF (a unique NotI restriction site has replaced a unique AccIrestriction site). The lacz gene is under the control of the syntheticlate promoter (LP1), and the BRSV G gene is under the control of thesynthetic late/early promoter (LP2EP2).

S-SPV-020 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 727-20.5 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-020. This virus was assayed forβ-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

S-SPV-020 was assayed for expression of BRSV-specific antigens using theBLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV.Bovine anti-BRSV FITC (Accurate Chemicals) was shown to reactspecifically with S-SPV-020 plaques and not with S-SPV-003 negativecontrol plaques. All S-SPV-020 observed plaques reacted with theantiserum indicating that the virus was stably expressing the BRSVforeign gene. The assays described here were carried out in ESK-4 cells,indicating that ESK-4 cells would be a suitable substrate for theproduction of SPV recombinant vaccines.

To confirm the expression of the BRSV G gene product, cells wereinfected with S-SPV-020 and samples of infected cell lysates weresubjected to SDS-polyacrylamide gel electrophoresis. The gel was blottedand analyzed using the WESTERN BLOTTING PROCEDURE. Bovine anti-BRSV FITC(Accurate Chemicals) was used to detect expression of BRSV specificproteins. The lysate from S-SPV-020 infected cells exhibited a band at36 kd which is the expected size of the non-glycosylated form of BRSV Gprotein and bands at 43 to 45 kd and 80 to 90 kd which are the expectedsize of glycosylated forms of the BRSV G protein.

Example 19B

S-SPV-029:

S-SPV-029 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene for bovinerespiratory syncytial virus (BRSV) F were inserted into the SPV 617-48.1ORF (a unique NotI restriction site has replaced a unique AccIrestriction site). The lacZ gene is under the control of the syntheticlate promoter (LP1), and the BRSV F gene is under the control of thesynthetic late/early promoter (LP2EP2).

S-SPV-029 was derived from S-SPV-O01 (Kasza Strain). This wasaccomplished utilizing the homology vector 727-20.10 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-029.

This virus was assayed for β-galactosidase expression, purity, andinsert stability by multiple passages monitored by the blue plaque assayas described in Materials and Methods. After the initial three rounds ofpurification, all plaques observed were blue indicating that the viruswas pure, stable, and expressing the foreign gene.

S-SPV-029 was assayed for expression of BRSV-specific antigens using theBLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV.Bovine anti-BRSV FITC (Accurate Chemicals) was shown to reactspecifically with S-SPV-029 plaques and not with S-SPV-003 negativecontrol plaques. All S-SPV-029 observed plaques reacted with theantiserum indicating that the virus was stably expressing the BRSVforeign gene. The assays described here were carried out in ESK-4 cells,indicating that ESK-4 cells would be a suitable substrate for theproduction of SPV recombinant vaccines.

Example 19C

S-SPV-030:

S-SPV-030 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene for bovinerespiratory syncytial virus (BRSV) N were inserted into the SPV 617-48.1ORF (a unique NotI restriction site has replaced a unique AccIrestriction site). The lacZ gene is under the control of the syntheticlate promoter (LP1), and the BRSV N gene is under the control of thesynthetic late/early promoter (LP2EP2).

S-SPV-030 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 713-55.37 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-030. This virus was assayed forβ-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

S-SPV-030 was assayed for expression of BRSV-specific antigens using theBLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV.Bovine anti-BRSV FITC (Accurate Chemicals) was shown to reactspecifically wit h S-SPV-030 plaques and not with S-SPV-003 negativecontrol plaques. All S-SPV-030 observed plaques reacted with theantiserum indicating that the virus was stably expressing the BRSVforeign gene. The assays described here were carried out in ESK-4 cells,indicating that ESK-4 cells would be a suitable substrate for theproduction of SPV recombinant vaccines.

To confirm the expression of the BRSV N gene product, cells wereinfected with SPV-030 and samples of infected cell lysates weresubjected to SDS-polyacrylamide gel electrophoresis. The gel was blottedand analyzed using the WESTERN BLOTTING PROCEDURE. Bovine anti-BRSV FITC(Accurate Chemicals) was used to detect expression of BRSV specificproteins. The lysate from S-SPV-030 infected cells exhibited a band at43 kd which is the expected size of the BRSV N protein.

Example 19D

S-SPV-028:

S-SPV-028 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene for bovineparainfluenza virus type 3 (BPI-3) F were inserted into the SPV 617-48.1ORF (a unique NotI restriction site has replaced a unique AccIrestriction site). The lacz gene is under the control of the syntheticlate promoter (LP1), and the BPI-3 F gene is under the control of thesynthetic late/early promoter (LP2EP2).

S-SPV-028 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 713-55.10 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-028. This virus was assayed forβ-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

S-SPV-028 was assayed for expression of BPI-3-specific antigens usingthe BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV.Bovine anti-BPI-3 FITC (Accurate Chemicals) was shown to reactspecifically with S-SPV-028 plaques and not with S-SPV-003 negativecontrol plaques. All S-SPV-028 observed plaques reacted with theantiserum indicating that the virus was stably expressing the BPI-3foreign gene. The assays described here were carried out in ESK-4 cells,indicating that ESK-4 cells would be a suitable substrate for theproduction of SPV recombinant vaccines. To confirm the expression of theBPI-3 F gene product, cells were infected with SPV-028 and samples ofinfected cell lysates were subjected to SDS-polyacrylamide gelelectrophoresis. The gel was blotted and analyzed using the WESTERNBLOTTING PROCEDURE. Bovine anti-BPI-3 FITC (Accurate Chemicals) was usedto detect expression of BPI-3 specific proteins. The lysate fromS-SPV-028 infected cells exhibited bands at 43, and 70 kd which is theexpected size of the BPI-3 F protein.

Example 19E

S-SPV-032:

S-SPV-032 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene for bovineviral diarrhea virus (BVDV) gp48 were inserted into the SPV 617-48.1 ORF(a unique NotI restriction site has replaced a unique AccI restrictionsite). The lacZ gene is under the control of the synthetic late promoter(LP1), and the BVDV gp48 gene is under the control of the syntheticlate/early promoter (LP2EP2).

S-SPV-032 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 727-78.1 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-032. This virus was assayed forβ-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

Example 19F

S-SPV-040:

S-SPV-040 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene for bovineviral diarrhea virus (BVDV) gp53 were inserted into the SPV 617-48.1 ORF(a unique NotI restriction site has replaced a unique AccI restrictionsite). The lacZ gene is under the control of the synthetic late promoter(LP1), and the BVDV gp53 gene is under the control of the syntheticlate/early promoter (LP2EP2).

S-SPV-040 is derived from S-SPV-001 (Kasza Strain). This is accomplishedutilizing the homology vector 738-96 (see Materials and Methods) andvirus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATINGRECOMBINANT SPV. The transfection stock is screened by the SCREEN FORRECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL AND CPRG ASSAYS).The final result of red plaque purification is the recombinant virusdesignated S-SPV-040. This virus is assayed for β-galactosidaseexpression, purity, and insert stability by multiple passages monitoredby the blue plaque assay as described in Materials and Methods. Afterthe initial three rounds of purification, all plaques observed are blueindicating that the virus is pure, stable, and expressing the foreigngene.

Example 19G

Shipping Fever Vaccine

Shipping fever or bovine respiratory disease (BRD) complex is manifestedas the result of a combination of infectious diseases of cattle andadditional stress related factors (52). Respiratory virus infectionsaugmented by pathophysiological effects of stress, alter thesusceptibility of cattle to Pasteurella organisms by a number ofmechanisms. Control of the viral infections that initiate BRD isessential to preventing the disease syndrome (53).

The major infectious disease pathogens that contribute to BRD includebut are not limited to infectious bovine rhinotracheitis virus (IBRV),parainfluenza virus type 3 (PI-3), bovine respiratory syncytial virus(BRSV), and Pasteurella haemolytica (53). Recombinant swinepox virusexpressing protective antigens to organisms causing BRD is useful as avaccine. S-SPV-020, S-SPV-029, S-SPV-030, S-SPV-032, and S-SPV-028 areuseful components of such a vaccine.

Example 20

Recombinant swinepox viruses S-SPV-031 and S-SPV-035 are useful as avaccine against human disease. S-SPV-031 expresses the core antigen ofhepatitis B virus. S-SPV-031 is useful against hepatitis B infection inhumans. S-SPV-035 expresses the cytokine, interleukin-2, and is usefulas an immune modulator to enhance an immune response in humans. WhenS-SPV-031 and S-SPV-035 are combined, a superior vaccine againsthepatitis B is produced.

Example 20A

S-SPV-031:

S-SPV-031 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene for Hepatitis BCore antigen were inserted into the SPV 617-48.1 ORF (a unique NotIrestriction site has replaced a unique AccI restriction site). The lacZgene is under the control of the synthetic late promoter (LP1), and theHepatitis B Core antigen gene is under the control of the syntheticearly/late promoter (EP1LP2).

S-SPV-031 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 727-67.18 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-031. This virus was assayed forβ-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

S-SPV-031 was assayed for expression of Hepatitis B Coreantigen-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENEEXPRESSION IN RECOMBINANT SPV. Rabbit antisera to Hepatitis B Coreantigen was shown to react specifically with S-SPV-031 plaques and notwith S-SPV-001 negative control plaques. All S-SPV-031 observed plaquesreacted with the antiserum indicating that the virus was stablyexpressing the Hepatitis B Core antigen gene. The assays described herewere carried out in ESK-4 cells, indicating that ESK-4 cells would be asuitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the Hepatitis B Core antigen gene product,cells were infected with SPV-031 and samples of infected cell lysateswere subjected to SDS-polyacrylamide gel electrophoresis. The gel wasblotted and analyzed using the WESTERN BLOTTING PROCEDURE. Rabbitantisera to Hepatitis B Core antigen was used to detect expression ofHepatitis B specific proteins. The lysate from S-SPV-031 infected cellsexhibited a band at 21 kd which is the expected size of the Hepatitis BCore antigen.

Example 20B

S-SPV-035:

S-SPV-035 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene for human IL-2were inserted into the SPV 617-48.1 ORF (a unique NotI restriction sitehas replaced a unique AccI restriction site). The lacZ gene is under thecontrol of the synthetic late promoter (LP1), and the human IL-2 gene isunder the control of the synthetic late/early promoter (LP2EP2).

S-SPV-035 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 741-84.14 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-035. This virus was assayed forβ-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

Example 21

Human vaccines using recombinant swinepox virus as a vector

Recombinant swinepox virus is useful as a vaccine against humandiseases. For example, human influenza virus is a rapidly evolving viruswhose neutralizing viral epitopes rapidly change. A useful recombinantswinepox vaccine is one in which the influenza virus neutralizingepitopes are quickly adapted by recombinant DNA techniques to protectagainst new strains of influenza virus. Human influenza virushemagglutinin (HN) and neuraminidase (NA) genes are cloned into theswinepox virus as described in CLONING OF EQUINE INFLUENZA VIRUSHEMAGGLUTININ AND NEURAMINIDASE GENES (See Materials and Methods andExample 17).

Recombinant swinepox virus is useful as a vaccine against other humandiseases when foreign antigens from the following diseases or diseaseorganisms are expressed in the swinepox virus vector: hepatitis B virussurface and core antigens, hepatitis C virus, human immunodeficiencyvirus, human herpesviruses, herpes simplex virus-1, herpes simplexvirus-2, human cytomegalovirus, Epstein-Barr virus, Varicella-Zostervirus, human herpesvirus-6, human herpesvirus-7, human influenza,measles virus, hantaan virus, pneumonia virus, rhinovirs, poliovirus,human respiratory syncytial virus, retrovirus, human T-cell leukemiavirus, rabies virus, mumps virus, malaria (Pasmodium falciparum),Bordetelia pertussis, Diptheria, Rickettsia prowazekii, Borrliabergdorferi, Tetanus toxoid, malignant tumor antigens.

Furthermore, S-SPV-035 (Example 20), when combined with swinepox virusinterleukin-2 is useful in enhancing immune response in humans.Additional cytokines, including but not limited to, interleukin-2,interleukin-6, interleukin-12, interferons, granulocyte-macrophagecolony stimulating factors, interleukin receptors from human and otheranimals when vectored into a non-essential site in the swinepox viralgenome, and subsequently expressed, have immune stimulating effects.

Recombinant swinepox virus express foreign genes in a human cell line.We demonstrated that S-SPV-003 (EP1LP2 promoter expressing the lacZ gen)expressed the lacZ gene in THP human monocyte cell lines by measuringβ-galactosidase activity. We did not observe any cytopathic effect ofswinepox virus on the THP human monocyte cells, indicating thatrecombinant swinepox virus can express foreign genes in a human cellline, but will not productively infect or replicated in the human cellline. We have demonstrated that swinepox virus replicates well in ESK-4cells (embryonic swine kidney) indicating that ESK-4 cells would be asuitable substrate for the production of SPV recombinant vaccines.

Example 22

Avian vaccines using recombinant swinepox virus as a vector

Example 22A

S-SPV-026

S-SPV-026 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene for infectiousbursal disease virus (IBDV) polyprotein were inserted into the SPV617-48.1 ORF (a unique NotI restriction site has replaced a unique AccIrestriction site). The lacZ gene is under the control of the syntheticlate promoter (LP1), and the IBDV polyprotein gene is under the controlof the synthetic early/late promoter (EP1LP2).

S-SPV-026 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 689-50.4 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-026. This virus was assayed forβ-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indication that the virus was pure, stable, andexpressing the foreign gene.

S-SPV-026 was assayed for expression of IBDV polyprotein-specificantigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION INRECOMBINANT SPV. Rat antisera to IBDV polyprotein were shown to reactspecifically with S-SPV-026 plaques and not with S-SPV-001 negativecontrol plaques. All S-SPV-026 observed plaques reacted with theantiserum indicating that the virus was stably expressing the IBDVpolyprotein gene. The assays described here were carried out in ESK-4cells, indicating that ESK-4 cells would be a suitable substrate for theproduction of SPV recombinant vaccines.

To confirm the expression of the IBDV polyprotein gene product, cellswere infected with SPV-026 and samples of infected cell lysates weresubjected to SDS-polyacrylamide gel electrophoresis. The gel was blottedand analyzed using the WESTERN BLOTTING PROCEDURE. Rat antisera to IBDVproteins VP2, VP3, and VP4 and monoclonal antibody R63 to IBDV VP2 wereused to detect expression of IBDV proteins. The lysate from S-SPV-026infected cells exhibited bands at 32 to 40 kd which is the expected sizeof the IBDV proteins.

Example 22B

S-SPV-027

S-SPV-027 is a swinepox virus that expresses at least two foreign genes.The gene for E. coli β-galactosidase (lacZ) and the gene for infectiousbursal disease virus (IBDV) VP2 (40 kd) were inserted into the SPV617-48.1 ORF (a unique NotI restriction site has replaced a unique AccIrestriction site). The lacZ gene is under the control of the syntheticlate promoter (LP1), and the IBDV VP2 gene is under the control of thesynthetic early/late promoter (EP1LP2).

S-SPV-027 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 689-50.7 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-027. This virus was assayed forβ-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved were blue indicating that the virus was pure, stable, andexpressing the foreign gene.

S-SPV-027 was assayed for expression of IBDV VP2-specific antigens usingthe BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV.Rat antisera to IBDV protein was shown to react specifically withS-SPV-027 plaques and not with S-SPV-001 negative control plaques. AllS-SPV-027 observed plaques reacted with the antiserum indicating thatthe virus was stably expressing the IBDV VP2 gene. The assays describedhere were carried out in ESK-4 cells, indicating that ESK-4 cells wouldbe a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the IBDV VP2 gene product, cells wereinfected with S-SPV-027 and samples of infected cell lysates weresubjected to SDS-polyacrylamide gel electrophoresis. The gel was blottedand analyzed using the WESTERN BLOTTING PROCEDURE. Rat antisera to IBDVprotein and monoclonal antibody R63 to IBDV VP2 were used to detectexpression of IBDV VP2 protein. The lysate from S-SPV-027 infected cellsexhibited a band at 40 kd which is the expected size of the IBDV VP2protein.

S-SPV-026 and S-SPV-027 are useful as vaccines against infectious bursaldisease in chickens and also as expression vectors for IBDV proteins.Recombinant swinepox virus is useful as a vaccine against other aviandisease when foreign antigens from the following diseases or diseaseorganisms are expressed in the swinepox virus vector: Marek's diseasevirus, infectious laryngotracheitis virus, Newcastle disease virus,infectious bronchitis virus, and chicken anemia virus.

Example 23

SPV-036:

S-SPV-036 is a swinepox virus that expresses at one foreign gene. Thegene for E. coli β-galactosidase (lacZ) was inserted into the SPV617-48.1 ORF (a unique NotI restriction site has replaced a unique AccIrestriction site). The lacZ gene is under the control of the humancytomegalovirus immediate early (HCMV IE) promoter.

S-SPV-036 was derived from S-SPV-001 (Kasza Strain). This wasaccomplished utilizing the homology vector 741-80.3 (see Materials andMethods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDUREFOR GENERATING RECOMBINANT SPV. The transfection stock was screened bythe SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase (BLUOGAL ANDCPRG ASSAYS). The final result of red plaque purification was therecombinant virus designated S-SPV-036. This virus is assayed forβ-galactosidase expression, purity, and insert stability by multiplepassages monitored by the blue plaque assay as described in Materialsand Methods. After the initial three rounds of purification, all plaquesobserved are blue indicating that the virus is pure, stable, andexpressing the foreign gene.

The expression of lacZ from the HCMV IE promoter provides a strongpromoter for expression of foreign genes in swinepox. S-SPV-036 is anovel and unexpected demonstration of a herpesvirus promoter drivingexpression of a foreign gene in a poxvirus. S-SPV-036 is useful informulating human vaccines, and recombinant swinepox virus is useful forthe expression of neutralizing antigens from human pathogens.Recombinant swinepox virus expressed foreign genes in a human cell lineas demonstrated by S-SPV-003 (EP1LP2) promoter expressing the lacZ gene)expressed β-galactosidase in THP human monocyte cell lines.

Recombinant swinepox virus expressed foreign genes in a human cell lineas demonstrated by s-SPV-003 (EP1LP2 promoter expressing the lacz gene)expressed β-galactosidase in THP human monocyte cell lines. THP humanmonocyte cells are useful for the production of recombinant swinepoxvirus as a human vaccine. Other cell lines in which swinepox virus willreplicate include, but are not limited to, Vero cells (monkey), ST cells(swine testicle), PK-15 (porcine kidney), and ESK-4 cells (embryonicswine kidney).

Example 24

Homology Vector 738-94.5

Homology Vector 738-94.5 is a swinepox virus vector that expresses oneforeign gene. The gene for E. coli β-galactosidase (lacZ) was insertedinto the the O1L open reading frame (SEQ ID NO. 115). The lacZ gene isunder the control of the O1L promoter. The homology vector 738-94.5contains a deletion of SPV DNA from nucleotides 1381 to 2133 (SEQ ID NO.113; FIG. 17) which deletes part of the O1L ORF.

The upstream SPV sequences were synthesized by polymerase chain reactionusing DNA primers 5'-GAAGCATGCCCGTTCTTATCAATAGTTTAGTCGAAAATA-3' (SEQ IDNO. 185) and 5'-CATAAGATCTGGCATTGTGTTATTATACTAACAAAAATAAG-3' (SEQ ID NO.186) to produce an 871 base pair fragment with BglII and SphI ends. TheO1L promoter is present on this fragment. The downstream SPV sequenceswere synthesized by polymerase chain reaction using DNA primers5'-CCGTAGTCGACAAAGATCGACTTATTAATATGTATGGGATT-3' (SEQ ID NO. 187) and5'-GCCTGAAGCTTCTAGTACAGTATTTACGACTTTTGAAAT-3' (SEQ ID NO. 188) toproduce an 1123 base pair fragment with SalI and HindIII ends. Arecombinant swinepox virus was derived utilizing homology vector738-94.5 and S-SPV-001 (Kasza strain) in the HOMOLOGOUS RECOMBINATIONPROCEDURE FOR GENERATING RECOMBINANT SPV. The transfection stock wasscreened by the SCREEN FOR RECOMBINANT SPV EXPRESSING β-galactosidase(BLUOGAL AND CPRG ASSAYS). The final result of red plaque purificationis the recombinant virus. This virus is assayed for β-galactosidaseexpression, purity, and insert stability by multiple passages monitoredby the blue plaque assay as described in Materials and Methods. Afterthe initial three rounds of purification, all plaques observed are blueindicating that the virus is pure, stable, and expressing the foreigngene. Recombinant swinepox viruses derived from homology vector 738-94.5are utilized as an expression vector to express foreign antigens and asa vaccine to raise a protective immune response in animals to foreigngenes expressed by the recombinant swinepox virus. Other promoters inaddition to the O1L promoter are inserted into the deleted regionincluding LP1, EP1LP2, LP2EP2, HCMV immediate early, and one or moreforei gn genes are expressed from these promoters.

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27. A. K. Robbins et al., Journal of Virology 58, 339-347 (1986).

28. A. K. Robbins et al., Journal of Virology 61, 2691-2701 (1987).

29. A. C. R. Samson, Journal of Virology 67, 1199-1203 (1986).

30. J. Sambrook, et al., Molecular Cloning A Laboratory Manual SecondEdition, Cold Spring Harbor Press, Cold Spring Harbor, N.Y. (1989).

31. Sheffy, et al., Proceedings 65th Annual Meeting of the United StatesLivestock Association 65, 347-353 (1961).

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33. J. Taylor, et al., Vaccine 9, 190-193, (1991).

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53. F. Fenner, et al., Veterinary Virology. Academic Press, Inc.,Orlando Fla., 183-202 (1987).

    __________________________________________________________________________    #             SEQUENCE LISTING                                                   - -  - - (1) GENERAL INFORMATION:                                             - -    (iii) NUMBER OF SEQUENCES: 188                                         - -  - - (2) INFORMATION FOR SEQ ID NO:1:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 599 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 202..597                                                        (D) OTHER INFORMATION: - #/partial                                                 /codon.sub.-- - #start= 202                                                   /function=- # "Potential eukaryotic transcriptional                          regulatory - #protein"                                                        /standard.sub.-- - #name= "515-85.1 ORF"                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                               - - AATGTATCCA GAGTTGTTGA ATGCCTTATC GTACCTAATA TTAATATAGA GT -             #TATTAACT     60                                                                 - - GAATAAGTAT ATATAAATGA TTGTTTTTAT AATGTTTGTT ATCGCATTTA GT -            #TTTGCTGT    120                                                                 - - ATGGTTATCA TATACATTTT TAAGGCCGTA TATGATAAAT GAAAATATAT AA -            #GCACTTAT    180                                                                 - - TTTTGTTAGT ATAATAACAC A ATG CCG TCG TAT ATG TAT - #CCG AAG AAC GCA          231                                                                                         - #      Met Pro Ser Tyr Met Tyr Pro - #Lys Asn Ala                           - #        1          - #     5             - #     10       - - AGA AAA GTA ATT TCA AAG ATT ATA TCA TTA CA - #A CTT GAT ATT AAA AAA          279                                                                       Arg Lys Val Ile Ser Lys Ile Ile Ser Leu Gl - #n Leu Asp Ile Lys Lys                            15 - #                 20 - #                 25              - -  CTT CCT AAA AAA TAT ATA AAT ACC ATG TTA - #GAA TTT GGT CTA CAT GGA         327                                                                        Leu Pro Lys Lys Tyr Ile Asn Thr Met Leu Gl - #u Phe Gly Leu His Gly                        30     - #             35     - #             40                  - - AAT CTA CCA GCT TGT ATG TAT AAA GAT GCC GT - #A TCA TAT GAT ATA AAT          375                                                                       Asn Leu Pro Ala Cys Met Tyr Lys Asp Ala Va - #l Ser Tyr Asp Ile Asn                    45         - #         50         - #         55                      - - AAT ATA AGA TTT TTA CCT TAT AAT TGT GTT AT - #G GTT AAA GAT TTA ATA          423                                                                       Asn Ile Arg Phe Leu Pro Tyr Asn Cys Val Me - #t Val Lys Asp Leu Ile                60             - #     65             - #     70                          - - AAT GTT ATA AAA TCA TCA TCT GTA ATA GAT AC - #T AGA TTA CAT CAA TCT          471                                                                       Asn Val Ile Lys Ser Ser Ser Val Ile Asp Th - #r Arg Leu His Gln Ser            75                 - # 80                 - # 85                 - # 90       - - GTA TTA AAA CAT CGT AGA GCG TTA ATA GAT TA - #C GGC GAT CAA GAC ATT          519                                                                       Val Leu Lys His Arg Arg Ala Leu Ile Asp Ty - #r Gly Asp Gln Asp Ile                            95 - #                100 - #                105              - - ATC ACT TTA ATG ATC ATT AAT AAG TTA CTA TC - #G ATA GAT GAT ATA TCC          567                                                                       Ile Thr Leu Met Ile Ile Asn Lys Leu Leu Se - #r Ile Asp Asp Ile Ser                       110      - #           115      - #           120                  - - TAT ATA TTA GAT AAA AAA ATA ATT CAT GTA AC - #                  - #             599                                                                    Tyr Ile Leu Asp Lys Lys Ile Ile His Val                                               125          - #       130                                             - -  - - (2) INFORMATION FOR SEQ ID NO:2:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 132 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                               - - Met Pro Ser Tyr Met Tyr Pro Lys Asn Ala Ar - #g Lys Val Ile Ser Lys        1               5 - #                 10 - #                 15              - - Ile Ile Ser Leu Gln Leu Asp Ile Lys Lys Le - #u Pro Lys Lys Tyr Ile                   20     - #             25     - #             30                  - - Asn Thr Met Leu Glu Phe Gly Leu His Gly As - #n Leu Pro Ala Cys Met               35         - #         40         - #         45                      - - Tyr Lys Asp Ala Val Ser Tyr Asp Ile Asn As - #n Ile Arg Phe Leu Pro           50             - #     55             - #     60                          - - Tyr Asn Cys Val Met Val Lys Asp Leu Ile As - #n Val Ile Lys Ser Ser       65                 - # 70                 - # 75                 - # 80       - - Ser Val Ile Asp Thr Arg Leu His Gln Ser Va - #l Leu Lys His Arg Arg                       85 - #                 90 - #                 95              - - Ala Leu Ile Asp Tyr Gly Asp Gln Asp Ile Il - #e Thr Leu Met Ile Ile                  100      - #           105      - #           110                  - - Asn Lys Leu Leu Ser Ile Asp Asp Ile Ser Ty - #r Ile Leu Asp Lys Lys              115          - #       120          - #       125                      - - Ile Ile His Val                                                              130                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:3:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 899 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 3..662                                                          (D) OTHER INFORMATION: - #/partial                                                 /codon.sub.-- - #start= 3                                                     /function=- # "Potential eukaryotic transcriptional                           regulatory - #protein"                                                        /standard.sub.-- - #name= "515-85.1 ORF"                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                               - - GA GAT ATT AAA TCA TGT AAA TGC TCG ATA TGT - # TCC GAC TCT ATA ACA            47                                                                          Asp Ile Lys Ser Cys Lys Cys Ser Ile - #Cys Ser Asp Ser Ile Thr                  1             - #  5                - #  10                - #  15        - - CAT CAT ATA TAT GAA ACA ACA TCA TGT ATA AA - #T TAT AAA TCT ACC GAT           95                                                                       His His Ile Tyr Glu Thr Thr Ser Cys Ile As - #n Tyr Lys Ser Thr Asp                            20 - #                 25 - #                 30              - - AAT GAT CTT ATG ATA GTA TTG TTC AAT CTA AC - #T AGA TAT TTA ATG CAT          143                                                                       Asn Asp Leu Met Ile Val Leu Phe Asn Leu Th - #r Arg Tyr Leu Met His                        35     - #             40     - #             45                  - - GGG ATG ATA CAT CCT AAT CTT ATA AGC GTA AA - #A GGA TGG GGT CCC CTT          191                                                                       Gly Met Ile His Pro Asn Leu Ile Ser Val Ly - #s Gly Trp Gly Pro Leu                    50         - #         55         - #         60                      - - ATT GGA TTA TTA ACG GGT GAT ATA GGT ATT AA - #T TTA AAA CTA TAT TCC          239                                                                        Ile Gly Leu Leu Thr Gly Asp Ile Gly Ile - #Asn Leu Lys Leu Tyr Ser                65             - #     70             - #     75                          - - ACC ATG AAT ATA AAT GGG CTA CGG TAT GGA GA - #T ATT ACG TTA TCT TCA          287                                                                       Thr Met Asn Ile Asn Gly Leu Arg Tyr Gly As - #p Ile Thr Leu Ser Ser            80                 - # 85                 - # 90                 - # 95       - - TAC GAT ATG AGT AAT AAA TTA GTC TCT ATT AT - #T AAT ACA CCC ATA TAT          335                                                                       Tyr Asp Met Ser Asn Lys Leu Val Ser Ile Il - #e Asn Thr Pro Ile Tyr                           100  - #               105  - #               110              - - GAG TTA ATA CCG TTT ACT ACA TGT TGT TCA CT - #C AAT GAA TAT TAT TCA          383                                                                       Glu Leu Ile Pro Phe Thr Thr Cys Cys Ser Le - #u Asn Glu Tyr Tyr Ser                       115      - #           120      - #           125                  - - AAA ATT GTG ATT TTA ATA AAT GTT ATT TTA GA - #A TAT ATG ATA TCT ATT          431                                                                       Lys Ile Val Ile Leu Ile Asn Val Ile Leu Gl - #u Tyr Met Ile Ser Ile                   130          - #       135          - #       140                      - - ATA TTA TAT AGA ATA TTG ATC GTA AAA AGA TT - #T AAT AAC ATT AAA GAA          479                                                                       Ile Leu Tyr Arg Ile Leu Ile Val Lys Arg Ph - #e Asn Asn Ile Lys Glu               145              - #   150              - #   155                          - - TTT ATT TCA AAA GTC GTA AAT ACT GTA CTA GA - #A TCA TCA GGC ATA TAT          527                                                                       Phe Ile Ser Lys Val Val Asn Thr Val Leu Gl - #u Ser Ser Gly Ile Tyr           160                 1 - #65                 1 - #70                 1 -      #75                                                                              - - TTT TGT CAG ATG CGT GTA CAT GAA CAA ATT GA - #A TTG GAA ATA GAT        GAG      575                                                                    Phe Cys Gln Met Arg Val His Glu Gln Ile Gl - #u Leu Glu Ile Asp Glu                          180  - #               185  - #               190              - - CTC ATT ATT AAT GGA TCT ATG CCT GTA CAG CT - #T ATG CAT TTA CTT CTA          623                                                                       Leu Ile Ile Asn Gly Ser Met Pro Val Gln Le - #u Met His Leu Leu Leu                       195      - #           200      - #           205                  - - AAG GTA GCT ACC ATA ATA TTA GAG GAA ATC AA - #A GAA ATA TAACGTATTT           672                                                                       Lys Val Ala Thr Ile Ile Leu Glu Glu Ile Ly - #s Glu Ile                               210          - #       215          - #       220                      - - TTTCTTTTAA ATAAATAAAA ATACTTTTTT TTTTAAACAA GGGGTGCTAC CT -             #TGTCTAAT    732                                                                 - - TGTATCTTGT ATTTTGGATC TGATGCAAGA TTATTAAATA ATCGTATGAA AA -            #AGTAGTAG    792                                                                 - - ATATAGTTTA TATCGTTACT GGACATGATA TTATGTTTAG TTAATTCTTC TT -            #TGGCATGA    852                                                                 - - ATTCTACACG TCGGANAAGG TAATGTATCT ATAATGGTAT AAAGCTT   - #                   899                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:4:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 220 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                               - - Asp Ile Lys Ser Cys Lys Cys Ser Ile Cys Se - #r Asp Ser Ile Thr His        1               5 - #                 10 - #                 15              - - His Ile Tyr Glu Thr Thr Ser Cys Ile Asn Ty - #r Lys Ser Thr Asp Asn                   20     - #             25     - #             30                  - - Asp Leu Met Ile Val Leu Phe Asn Leu Thr Ar - #g Tyr Leu Met His Gly               35         - #         40         - #         45                      - - Met Ile His Pro Asn Leu Ile Ser Val Lys Gl - #y Trp Gly Pro Leu Ile           50             - #     55             - #     60                          - - Gly Leu Leu Thr Gly Asp Ile Gly Ile Asn Le - #u Lys Leu Tyr Ser Thr       65                 - # 70                 - # 75                 - # 80       - - Met Asn Ile Asn Gly Leu Arg Tyr Gly Asp Il - #e Thr Leu Ser Ser Tyr                       85 - #                 90 - #                 95              - - Asp Met Ser Asn Lys Leu Val Ser Ile Ile As - #n Thr Pro Ile Tyr Glu                  100      - #           105      - #           110                  - - Leu Ile Pro Phe Thr Thr Cys Cys Ser Leu As - #n Glu Tyr Tyr Ser Lys              115          - #       120          - #       125                      - - Ile Val Ile Leu Ile Asn Val Ile Leu Glu Ty - #r Met Ile Ser Ile Ile          130              - #   135              - #   140                          - - Leu Tyr Arg Ile Leu Ile Val Lys Arg Phe As - #n Asn Ile Lys Glu Phe      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Ile Ser Lys Val Val Asn Thr Val Leu Glu Se - #r Ser Gly Ile Tyr        Phe                                                                                             165  - #               170  - #               175             - - Cys Gln Met Arg Val His Glu Gln Ile Glu Le - #u Glu Ile Asp Glu Leu                  180      - #           185      - #           190                  - - Ile Ile Asn Gly Ser Met Pro Val Gln Leu Me - #t His Leu Leu Leu Lys              195          - #       200          - #       205                      - - Val Ala Thr Ile Ile Leu Glu Glu Ile Lys Gl - #u Ile                          210              - #   215              - #   220                          - -  - - (2) INFORMATION FOR SEQ ID NO:5:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 129 amino - #acids                                                (B) TYPE: amino acid                                                          (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: peptide                                           - -    (iii) HYPOTHETICAL: YES                                                - -     (iv) ANTI-SENSE: NO                                                   - -      (v) FRAGMENT TYPE: N-terminal                                        - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Vaccinia - #virus                                               (B) STRAIN: Copenhagen                                               - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                               - - Met Phe Met Tyr Pro Glu Phe Ala Arg Lys Al - #a Leu Ser Lys Leu Ile      1               5   - #                10  - #                15               - - Ser Lys Lys Leu Asn Ile Glu Lys Val Ser Se - #r Lys His Gln Leu Val                  20      - #            25      - #            30                   - - Leu Leu Asp Tyr Gly Leu His Gly Leu Leu Pr - #o Lys Ser Leu Tyr Leu              35          - #        40          - #        45                       - - Glu Ala Ile Asn Ser Asp Ile Leu Asn Val Ar - #g Phe Phe Pro Pro Glu          50              - #    55              - #    60                           - - Ile Ile Asn Val Thr Asp Ile Val Lys Ala Le - #u Gln Asn Ser Cys Arg      65                  - #70                  - #75                  - #80        - - Val Asp Glu Tyr Leu Lys Ala Val Ser Leu Ty - #r His Lys Asn Ser Leu                      85  - #                90  - #                95               - - Met Val Ser Gly Pro Asn Val Val Lys Leu Me - #t Ile Glu Tyr Asn Leu                  100      - #           105      - #           110                  - - Leu Thr His Ser Asp Leu Glu Trp Leu Ile As - #n Glu Asn Val Val Lys              115          - #       120          - #       125                      - - Ala                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:6:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 132 amino - #acids                                                (B) TYPE: amino acid                                                          (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: peptide                                           - -    (iii) HYPOTHETICAL: YES                                                - -     (iv) ANTI-SENSE: NO                                                   - -      (v) FRAGMENT TYPE: N-terminal                                        - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                    - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                               - - Met Pro Ser Tyr Met Tyr Pro Lys Asn Ala Ar - #g Lys Val Ile Ser Lys      1               5   - #                10  - #                15               - - Ile Ile Ser Leu Gln Leu Asp Ile Lys Lys Le - #u Pro Lys Lys Tyr Ile                  20      - #            25      - #            30                   - - Asn Thr Met Leu Glu Phe Gly Leu His Gly As - #n Leu Pro Ala Cys Met              35          - #        40          - #        45                       - - Tyr Lys Asp Ala Val Ser Tyr Asp Ile Asn As - #n Ile Arg Phe Leu Pro          50              - #    55              - #    60                           - - Tyr Asn Cys Val Met Val Lys Asp Leu Ile As - #n Val Ile Lys Ser Ser      65                  - #70                  - #75                  - #80        - - Ser Val Ile Asp Thr Arg Leu His Gln Ser Va - #l Leu Lys His Arg Arg                      85  - #                90  - #                95               - - Ala Leu Ile Asp Tyr Gly Asp Gln Asp Ile Il - #e Thr Leu Met Ile Ile                  100      - #           105      - #           110                  - - Asn Lys Leu Leu Ser Ile Asp Asp Ile Ser Ty - #r Ile Leu Asp Lys Lys              115          - #       120          - #       125                      - - Ile Ile His Val                                                              130                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:7:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 101 amino - #acids                                                (B) TYPE: amino acid                                                          (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: peptide                                           - -    (iii) HYPOTHETICAL: YES                                                - -     (iv) ANTI-SENSE: NO                                                   - -      (v) FRAGMENT TYPE: C-terminal                                        - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Vaccinia - #virus                                               (B) STRAIN: Copenhagen                                               - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                               - - Val Leu Asn Asp Gln Tyr Ala Lys Ile Val Il - #e Phe Phe Asn Thr Ile      1               5   - #                10  - #                15               - -  Ile Glu Tyr Ile Ile Ala Thr Ile Tyr Tyr - #Arg Leu Thr Val Leu Asn                  20      - #            25      - #            30                   - - Asn Tyr Thr Asn Val Lys His Phe Val Ser Ly - #s Val Leu His Thr Val              35          - #        40          - #        45                       - - Met Glu Ala Cys Gly Val Leu Phe Ser Tyr Il - #e Lys Val Asn Asp Lys          50              - #    55              - #    60                           - - Ile Glu His Glu Leu Glu Glu Met Val Asp Ly - #s Gly Thr Val Pro Ser      65                  - #70                  - #75                  - #80        - - Tyr Leu Tyr His Leu Ser Ile Asn Val Ile Se - #r Ile Ile Leu Asp Asp                      85  - #                90  - #                95               - - Ile Asn Gly Thr Arg                                                                  100                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:8:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 100 amino - #acids                                                (B) TYPE: amino acid                                                          (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: peptide                                           - -    (iii) HYPOTHETICAL: YES                                                - -     (iv) ANTI-SENSE: NO                                                   - -      (v) FRAGMENT TYPE: C-terminal                                        - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                    - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                               - - Ser Leu Asn Glu Tyr Tyr Ser Lys Ile Val Il - #e Leu Ile Asn Val Ile      1               5   - #                10  - #                15               - - Leu Glu Tyr Met Ile Ser Ile Ile Leu Tyr Ar - #g Ile Leu Ile Val Lys                  20      - #            25      - #            30                   - - Arg Phe Asn Asn Ile Lys Glu Phe Ile Ser Ly - #s Val Val Asn Thr Val              35          - #        40          - #        45                       - - Leu Glu Ser Ser Gly Ile Tyr Phe Cys Gln Me - #t Arg Val His Glu Gln          50              - #    55              - #    60                           - - Ile Glu Leu Glu Ile Asp Glu Leu Ile Ile As - #n Gly Ser Met Pro Val      65                  - #70                  - #75                  - #80        - - Gln Leu Met His Leu Leu Leu Lys Val Ala Th - #r Ile Ile Leu Glu Glu                      85  - #                90  - #                95               - - Ile Lys Glu Ile                                                                      100                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:9:                                     - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 102 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: circular                                               - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Plasmid                                                - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 520-17.5 (Ju - #nction A)                                 - -      (x) PUBLICATION INFORMATION:                                                  (A) AUTHORS: Ferrari, F - #ranco A                                                 Trach, Ka - #thleen                                                           Hoch, Jam - #es A                                                        (B) TITLE: Sequence Ana - #lysis of the spo0B Locus Revels a                       Polycistroni - #c Transcription Unit                                     (C) JOURNAL: J. Bacteri - #ol.                                                (D) VOLUME: 161                                                               (E) ISSUE: 2                                                                  (F) PAGES: 556-562                                                            (G) DATE: Feb.-1985                                                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                               - - CACATACGAT TTAGGTGACA CTATAGAATA CAAGCTTTAT ACCATTATAG AT -             #ACATTACC     60                                                                 - - TTGTCCGACG TGTAGAATTC ATGCCAAAGA AGAATTAACT AA    - #                      - # 102                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:10:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 102 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: circular                                               - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Plasmid                                                - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 520-17.5 (Ju - #nction B)                                 - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 85..99                                                          (D) OTHER INFORMATION: - #/codon.sub.-- start= 85                                  /function=- # "Translational start of hybrid protein"                         /product=- # "N-terminal peptide"                                             /number= - #1                                                                 /standard.sub.-- - #name= "Translation of synthetic DNA                       sequence"                                                       - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 100..102                                                        (C) IDENTIFICATION METHOD: - # experimental                                   (D) OTHER INFORMATION: - #/partial                                                 /codon.sub.-- - #start= 100                                                   /function=- # "marker enzyme"                                                 /product=- # "Beta-Galactosidase"                                             /evidence=- # EXPERIMENTAL                                                    /gene= - #"lacZ"                                                              /number= - #2                                                                 /citation=- # ([1])                                             - -      (x) PUBLICATION INFORMATION:                                                  (A) AUTHORS: Ferrari, F - #ranco A                                                 Trach, Ka - #thleen                                                           Hoch, Jam - #es A                                                        (B) TITLE: Seqquence An - #alysis of the spo0B Locus Reveals                       a Polycis - #tronic Transcription Unit                                   (C) JOURNAL: J. Bacteri - #ol.                                                (D) VOLUME: 161                                                               (E) ISSUE: 2                                                                  (F) PAGES: 556-562                                                            (G) DATE: Feb.-1985                                                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                              - - GTAGTCGACT CTAGAAAAAA TTGAAAAACT ATTCTAATTT ATTGCACGGA GA -             #TCTTTTTT     60                                                                 - - TTTTTTTTTT TTTTTGGCAT ATAA ATG AAT TCG GAT CCC G - #TC                      - # 102                                                                                     - #         Met Asn Ser Asp Pro - #Val                                        - #           1       - #        5   1                       - -  - - (2) INFORMATION FOR SEQ ID NO:11:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 5 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                              - - Met Asn Ser Asp Pro                                                        1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:12:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                              - - Val                                                                        1                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:13:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 103 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: circular                                               - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Plasmid                                                - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 520-17.5 (Ju - #nction C)                                 - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..72                                                           (C) IDENTIFICATION METHOD: - # experimental                                   (D) OTHER INFORMATION: - #/partial                                                 /codon.sub.-- - #start= 1                                                     /function=- # "marker enzyme"                                                 /product=- # "Beta-galactosidase"                                             /evidence=- # EXPERIMENTAL                                                    /gene= - #"lacZ"                                                              /number= - #1                                                                 /citation=- # ([1])                                             - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 73..78                                                          (C) IDENTIFICATION METHOD: - # experimental                                   (D) OTHER INFORMATION: - #/codon.sub.-- start= 73                                  /function=- # "Translational finish of hybrid                                 protein"                                                                      /product=- # "C-terminal peptide"                                             /evidence=- # EXPERIMENTAL                                                    /number= - #2                                                                 /standard.sub.-- - #name= "Translation of synthetic DNA                       sequence"                                                       - -      (x) PUBLICATION INFORMATION:                                                  (A) AUTHORS: Ferrari, F - #ranco A                                                 Trach, Ka - #thleen                                                           Hoch, Jam - #es A                                                        (B) TITLE: Seqquence An - #alysis of the spo0B Locus Reveals                       a Polycis - #tronic Transcription Unit                                   (C) JOURNAL: J. Bacteri - #ol.                                                (D) VOLUME: 161                                                               (E) ISSUE: 2                                                                  (F) PAGES: 556-562                                                            (G) DATE: Feb.-1985                                                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                              - - AGC CCG TCA GTA TCG GCG GAA ATC CAG CTG AG - #C GCC GGT CGC TAC CAT           48                                                                       Ser Pro Ser Val Ser Ala Glu Ile Gln Leu Se - #r Ala Gly Arg Tyr His             1               5 - #                 10 - #                 15              - - TAC CAG TTG GTC TGG TGT CAA AAA GAT CCA TA - #ATTAATTA ACCCGGGTCG             98                                                                       Tyr Gln Leu Val Trp Cys Gln Lys Asp Pro                                                    20     - #              1                                         - - AAGAC                 - #                  - #                  - #               103                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:14:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                              - - Ser Pro Ser Val Ser Ala Glu Ile Gln Leu Se - #r Ala Gly Arg Tyr His        1               5 - #                 10 - #                 15              - - Tyr Gln Leu Val Trp Cys Gln Lys                                                       20                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:15:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                              - - Asp Pro                                                                    1                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:16:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 48 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: circular                                               - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Plasmid                                                - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 520-17.5 (Ju - #nction D)                                 - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                              - - AGATCCCCGG GCGAGCTCGA ATTCGTAATC ATGGTCATAG CTGTTTCC  - #                    48                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:17:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 57 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: circular                                               - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Plasmid                                                - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 538-46.26 (J - #unction A)                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                              - - CACATACGAT TTAGGTGACA CTATAGAATA CAAGCTTTAT ACCATTATAG AT - #ACATT            57                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:18:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 102 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: circular                                               - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Plasmid                                                - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 538-46.16 (J - #unction B)                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 91..102                                                         (C) IDENTIFICATION METHOD: - # experimental                                   (D) OTHER INFORMATION: - #/partial                                                 /codon.sub.-- - #start= 91                                                    /function=- # "marker enzyme"                                                 /product=- # "Beta-Galactosidase"                                             /evidence=- # EXPERIMENTAL                                                    /gene= - #"lacZ"                                                              /number= - #2                                                                 /citation=- # ([1])                                             - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 76..90                                                          (D) OTHER INFORMATION: - #/partial                                                 /codon.sub.-- - #start= 76                                                    /function=- # "Translational start of hybrid protein"                         /product=- # "N-terminal peptide"                                             /number= - #1                                                                 /standard.sub.-- - #name= "Translation of synthetic DNA                       sequence"                                                       - -      (x) PUBLICATION INFORMATION:                                                  (A) AUTHORS: Ferrari, F - #ranco A                                                 Trach, Ka - #thleen                                                           Hoch, Jam - #es A                                                        (B) TITLE: Seqquence An - #alysis of the spo0B Locus Reveals                       a Polycis - #tronic Transcription Unit                                   (C) JOURNAL: J. Bacteri - #ol.                                                (D) VOLUME: 161                                                               (E) ISSUE: 2                                                                  (F) PAGES: 556-562                                                            (G) DATE: Feb.-1985                                                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                              - - AAGCTGGTAG ATTTCCATGT AGGGCCGCCT GCAGGTCGAC TCTAGAATTT CA -             #TTTTGTTT     60                                                                 - - TTTTCTATGC TATAA ATG AAT TCG GAT CCC GTC GTT - #TTA CAA                    - # 102                                                                                     Met - #Asn Ser Asp Pro Val Val Leu Gln                                         - # 1               5  - # 1                                 - -  - - (2) INFORMATION FOR SEQ ID NO:19:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 5 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                              - - Met Asn Ser Asp Pro                                                        1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:20:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 4 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                              - - Val Val Leu Gln                                                            1                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:21:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 206 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: circular                                               - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Plasmid                                                - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 538-46.16 (J - #unction C)                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..63                                                           (C) IDENTIFICATION METHOD: - # experimental                                   (D) OTHER INFORMATION: - #/partial                                                 /codon.sub.-- - #start= 1                                                     /function=- # "marker enzyme"                                                 /product=- # "Beta-galactosidase"                                             /evidence=- # EXPERIMENTAL                                                    /number= - #1                                                                 /citation=- # ([1])                                             - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 64..69                                                          (C) IDENTIFICATION METHOD: - # experimental                                   (D) OTHER INFORMATION: - #/codon.sub.-- start= 64                                  /function=- # "Translational finish of hybrid                                 protein"                                                                      /product=- # "C-terminal peptide"                                             /evidence=- # EXPERIMENTAL                                                    /standard.sub.-- - #name= "Translation of synthetic DNA                       sequence"                                                       - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 177..185                                                        (C) IDENTIFICATION METHOD: - # experimental                                   (D) OTHER INFORMATION: - #/codon.sub.-- start= 177                                 /function=- # "Translational start of hybrid protein"                         /product=- # "N-terminal peptide"                                             /evidence=- # EXPERIMENTAL                                                    /standard.sub.-- - #name= "Translation of synthetic DNA                       sequence"                                                       - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 186..206                                                        (C) IDENTIFICATION METHOD: - # experimental                                   (D) OTHER INFORMATION: - #/partial                                                 /codon.sub.-- - #start= 186                                                   /function=- # "glycoprotein"                                                  /product=- # "PRV gp50"                                                       /evidence=- # EXPERIMENTAL                                                    /gene= - #"gp50"                                                              /number= - #3                                                                 /citation=- # ([2])                                             - -      (x) PUBLICATION INFORMATION:                                                  (A) AUTHORS: Ferrari, F - #ranco A                                                 Trach, Ka - #thleen                                                           Hoch, Jam - #es A                                                        (B) TITLE: Seqquence An - #alysis of the spo0B Locus Reveals                       a Polycis - #tronic Transcription Unit                                   (C) JOURNAL: J. Bacteri - #ol.                                                (D) VOLUME: 161                                                               (E) ISSUE: 2                                                                  (F) PAGES: 556-562                                                            (G) DATE: Feb.-1985                                                  - -      (x) PUBLICATION INFORMATION:                                                  (A) AUTHORS: Petrovskis, - #Erik A                                                 Timmins, - #James G                                                           Armentrout, - #Marty A                                                        Marchioli, - #Carmine C                                                       Jr. Yancy - #, Robert J                                                       Post, Leo - #nard E                                                      (B) TITLE: DNA Sequence - # of the Gene for Pseudorabies                           Virus gp5 - #0, a Glycoprotein without N-Linked                               Glycosylatio - #n                                                        (C) JOURNAL: J. Virol.                                                        (D) VOLUME: 59                                                                (E) ISSUE: 2                                                                  (F) PAGES: 216-223                                                            (G) DATE: Aug.-1986                                                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                              - - GTA TCG GCG GAA ATC CAG CTG AGC GCC GGT CG - #C TAC CAT TAC CAG TTG           48                                                                       Val Ser Ala Glu Ile Gln Leu Ser Ala Gly Ar - #g Tyr His Tyr Gln Leu             1               5 - #                 10 - #                 15              - - GTC TGG TGT CAA AAA GAT CCA TAATTAATTA ACCCGGCCG - #C CTGCAGGTCG              99                                                                       Val Trp Cys Gln Lys Asp Pro                                                                20     - #  1                                                     - - ACTCTAGAAA AAATTGAAAA ACTATTCTAA TTTATTGCAC GGAGATCTTT TT -             #TTTTTTTT    159                                                                 - - TTTTTTTTGG CATATAA ATG AAT TCG CTC GCA GCG CTA - #TTG GCG GCG               206                                                                                         - # Met Asn Ser Leu Ala Ala Leu Leu Ala Ala                                   - #   1           1    - #           5                       - -  - - (2) INFORMATION FOR SEQ ID NO:22:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 21 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                              - - Val Ser Ala Glu Ile Gln Leu Ser Ala Gly Ar - #g Tyr His Tyr Gln Leu        1               5 - #                 10 - #                 15              - - Val Trp Cys Gln Lys                                                                   20                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:23:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                              - - Asp Pro                                                                    1                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:24:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 3 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                              - - Met Asn Ser                                                                1                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:25:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                              - - Leu Ala Ala Leu Leu Ala Ala                                               1               5                                                             - -  - - (2) INFORMATION FOR SEQ ID NO:26:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 101 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: circular                                               - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Plasmid                                                - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 538-46.16 (J - #unction D)                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..15                                                           (D) OTHER INFORMATION: - #/partial                                                 /codon.sub.-- - #start= 1                                                     /function=- # "glycoprotein"                                                  /product=- # "PRV gp63"                                                       /gene= - #"gp63"                                                              /number= - #1                                                                 /citation=- # ([1])                                             - -      (x) PUBLICATION INFORMATION:                                                  (A) AUTHORS: Petrovskis, - #Erik A                                                 Timmins, - #James G                                                           Post, Len - #oard E                                                      (B) TITLE: Use of La - #mbda-gt11 To Isolate Genes for two                         Pseudorabies - # Virus Glycoproteins with homology to                         Herpes Si - #mplex Virus and Varicella-Zoster Virus                           Glycoprotein - #s                                                        (C) JOURNAL: J. Virol.                                                        (D) VOLUME: 60                                                                (E) ISSUE: 1                                                                  (F) PAGES: 185-193                                                            (G) DATE: Oct.-1986                                                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                              - - CGC GTG CAC CAC GAG GGACTCTAGA GGATCCATAA TTAATTAAT - #T AATTTTTATC           55                                                                       Arg Val His His Glu                                                             1               5                                                            - - CCGGGTCGAC CTGCAGGCGG CCGGGTCGAC CTGCAGGCGG CCAGAC   - #                    101                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:27:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 5 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                              - - Arg Val His His Glu                                                        1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:28:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 57 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: circular                                               - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Plasmid                                                - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 538-46.16 (J - #unction E)                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                              - - AGATCCCCGG GCGAGCTCGA ATTCGTAATC ATGGTCATAG CTGTTTCCTG TG - #TGAAA            57                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:29:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1907 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: cDNA to mRNA                                      - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Newcastle - #disease virus                                      (B) STRAIN: B1                                                       - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 137-23.803 ( - #PSY1142)                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  50%                                                        (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 92..1822                                                        (D) OTHER INFORMATION: - #/codon.sub.-- start= 92                                  /product=- # "NDV heamagglutinin-Neuraminidase"                               /gene= - #"HN"                                                                /number= - #1                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                              - - ACGGGTAGAA CGGTAAGAGA GGCCGCCCCT CAATTGCGAG CCAGACTTCA CA -             #ACCTCCGT     60                                                                 - - TCTACCGCTT CACCGACAAC AGTCCTCAAT C ATG GAC CGC GCC - #GTT AGC CAA           112                                                                                         - #                 Met - # Asp Arg Ala Val Ser Gln                           - #                  - # 1               5                   - - GTT GCG TTA GAG AAT GAT GAA AGA GAG GCA AA - #A AAT ACA TGG CGC TTG          160                                                                       Val Ala Leu Glu Asn Asp Glu Arg Glu Ala Ly - #s Asn Thr Trp Arg Leu                    10         - #         15         - #         20                      - - ATA TTC CGG ATT GCA ATC TTA TTC TTA ACA GT - #A GTG ACC TTG GCT ATA          208                                                                        Ile Phe Arg Ile Ala Ile Leu Phe Leu Thr - #Val Val Thr Leu Ala Ile                25             - #     30             - #     35                          - - TCT GTA GCC TCC CTT TTA TAT AGC ATG GGG GC - #T AGC ACA CCT AGC GAT          256                                                                       Ser Val Ala Ser Leu Leu Tyr Ser Met Gly Al - #a Ser Thr Pro Ser Asp            40                 - # 45                 - # 50                 - # 55       - - CTT GTA GGC ATA CCG ACT AGG ATT TCC AGG GC - #A GAA GAA AAG ATT ACA          304                                                                       Leu Val Gly Ile Pro Thr Arg Ile Ser Arg Al - #a Glu Glu Lys Ile Thr                            60 - #                 65 - #                 70              - - TCT ACA CTT GGT TCC AAT CAA GAT GTA GTA GA - #T AGG ATA TAT AAG CAA          352                                                                       Ser Thr Leu Gly Ser Asn Gln Asp Val Val As - #p Arg Ile Tyr Lys Gln                        75     - #             80     - #             85                  - - GTG GCC CTT GAG TCT CCA TTG GCA TTG TTA AA - #T ACT GAG ACC ACA ATT          400                                                                       Val Ala Leu Glu Ser Pro Leu Ala Leu Leu As - #n Thr Glu Thr Thr Ile                    90         - #         95         - #        100                      - - ATG AAC GCA ATA ACA TCT CTC TCT TAT CAG AT - #T AAT GGA GCT GCA AAC          448                                                                       Met Asn Ala Ile Thr Ser Leu Ser Tyr Gln Il - #e Asn Gly Ala Ala Asn               105              - #   110              - #   115                          - - AAC AGC GGG TGG GGG GCA CCT ATT CAT GAC CC - #A GAT TAT ATA GGG GGG          496                                                                       Asn Ser Gly Trp Gly Ala Pro Ile His Asp Pr - #o Asp Tyr Ile Gly Gly           120                 1 - #25                 1 - #30                 1 -      #35                                                                              - - ATA GGC AAA GAA CTC ATT GTA GAT GAT GCT AG - #T GAT GTC ACA TCA        TTC      544                                                                    Ile Gly Lys Glu Leu Ile Val Asp Asp Ala Se - #r Asp Val Thr Ser Phe                          140  - #               145  - #               150              - - TAT CCC TCT GCA TTT CAA GAA CAT CTG AAT TT - #T ATC CCG GCG CCT ACT          592                                                                       Tyr Pro Ser Ala Phe Gln Glu His Leu Asn Ph - #e Ile Pro Ala Pro Thr                       155      - #           160      - #           165                  - - ACA GGA TCA GGT TGC ACT CGA ATA CCC TCA TT - #T GAC ATG AGT GCT ACC          640                                                                       Thr Gly Ser Gly Cys Thr Arg Ile Pro Ser Ph - #e Asp Met Ser Ala Thr                   170          - #       175          - #       180                      - - CAT TAC TGC TAC ACC CAT AAT GTA ATA TTG TC - #T GGA TGC AGA GAT CAC          688                                                                       His Tyr Cys Tyr Thr His Asn Val Ile Leu Se - #r Gly Cys Arg Asp His               185              - #   190              - #   195                          - - TCA CAC TCA CAT CAG TAT TTA GCA CTT GGT GT - #G CTC CGG ACA TCT GCA          736                                                                       Ser His Ser His Gln Tyr Leu Ala Leu Gly Va - #l Leu Arg Thr Ser Ala           200                 2 - #05                 2 - #10                 2 -      #15                                                                              - - ACA GGG AGG GTA TTC TTT TCT ACT CTG CGT TC - #C ATC AAC CTG GAC        GAC      784                                                                    Thr Gly Arg Val Phe Phe Ser Thr Leu Arg Se - #r Ile Asn Leu Asp Asp                          220  - #               225  - #               230              - - ACC CAA AAT CGG AAG TCT TGC AGT GTG AGT GC - #A ACT CCC CTG GGT TGT          832                                                                       Thr Gln Asn Arg Lys Ser Cys Ser Val Ser Al - #a Thr Pro Leu Gly Cys                       235      - #           240      - #           245                  - - GAT ATG CTG TGC TCG AAA GCC ACG GAG ACA GA - #G GAA GAA GAT TAT AAC          880                                                                       Asp Met Leu Cys Ser Lys Ala Thr Glu Thr Gl - #u Glu Glu Asp Tyr Asn                   250          - #       255          - #       260                      - - TCA GCT GTC CCT ACG CGG ATG GTA CAT GGG AG - #G TTA GGG TTC GAC GGC          928                                                                       Ser Ala Val Pro Thr Arg Met Val His Gly Ar - #g Leu Gly Phe Asp Gly               265              - #   270              - #   275                          - - CAA TAT CAC GAA AAG GAC CTA GAT GTC ACA AC - #A TTA TTC GGG GAC TGG          976                                                                       Gln Tyr His Glu Lys Asp Leu Asp Val Thr Th - #r Leu Phe Gly Asp Trp           280                 2 - #85                 2 - #90                 2 -      #95                                                                              - - GTG GCC AAC TAC CCA GGA GTA GGG GGT GGA TC - #T TTT ATT GAC AGC        CGC     1024                                                                    Val Ala Asn Tyr Pro Gly Val Gly Gly Gly Se - #r Phe Ile Asp Ser Arg                          300  - #               305  - #               310              - - GTG TGG TTC TCA GTC TAC GGA GGG TTA AAA CC - #C AAT ACA CCC AGT GAC         1072                                                                       Val Trp Phe Ser Val Tyr Gly Gly Leu Lys Pr - #o Asn Thr Pro Ser Asp                       315      - #           320      - #           325                  - - ACT GTA CAG GAA GGG AAA TAT GTG ATA TAC AA - #G CGA TAC AAT GAC ACA         1120                                                                       Thr Val Gln Glu Gly Lys Tyr Val Ile Tyr Ly - #s Arg Tyr Asn Asp Thr                   330          - #       335          - #       340                      - - TGC CCA GAT GAG CAA GAC TAC CAG ATT CGA AT - #G GCC AAG TCT TCG TAT         1168                                                                       Cys Pro Asp Glu Gln Asp Tyr Gln Ile Arg Me - #t Ala Lys Ser Ser Tyr               345              - #   350              - #   355                          - - AAG CCT GGA CGG TTT GGT GGG AAA CGC ATA CA - #G CAG GCT ATC TTA TCT         1216                                                                       Lys Pro Gly Arg Phe Gly Gly Lys Arg Ile Gl - #n Gln Ala Ile Leu Ser           360                 3 - #65                 3 - #70                 3 -      #75                                                                              - - ATC AAA GTG TCA ACA TCC TTA GGC GAA GAC CC - #G GTA CTG ACT GTA        CCG     1264                                                                     Ile Lys Val Ser Thr Ser Leu Gly Glu Asp - #Pro Val Leu Thr Val Pro                          380  - #               385  - #               390              - - CCC AAC ACA GTC ACA CTC ATG GGG GCC GAA GG - #C AGA ATT CTC ACA GTA         1312                                                                       Pro Asn Thr Val Thr Leu Met Gly Ala Glu Gl - #y Arg Ile Leu Thr Val                       395      - #           400      - #           405                  - - GGG ACA TCC CAT TTC TTG TAT CAG CGA GGG TC - #A TCA TAC TTC TCT CCC         1360                                                                       Gly Thr Ser His Phe Leu Tyr Gln Arg Gly Se - #r Ser Tyr Phe Ser Pro                   410          - #       415          - #       420                      - - GCG TTA TTA TAT CCT ATG ACA GTC AGC AAC AA - #A ACA GCC ACT CTT CAT         1408                                                                       Ala Leu Leu Tyr Pro Met Thr Val Ser Asn Ly - #s Thr Ala Thr Leu His               425              - #   430              - #   435                          - - AGT CCT TAT ACA TTC AAT GCC TTC ACT CGG CC - #A GGT AGT ATC CCT TGC         1456                                                                       Ser Pro Tyr Thr Phe Asn Ala Phe Thr Arg Pr - #o Gly Ser Ile Pro Cys           440                 4 - #45                 4 - #50                 4 -      #55                                                                              - - CAG GCT TCA GCA AGA TGC CCC AAC TCA TGT GT - #T ACT GGA GTC TAT        ACA     1504                                                                    Gln Ala Ser Ala Arg Cys Pro Asn Ser Cys Va - #l Thr Gly Val Tyr Thr                          460  - #               465  - #               470              - - GAT CCA TAT CCC CTA ATC TTC TAT AGA AAC CA - #C ACC TTG CGA GGG GTA         1552                                                                       Asp Pro Tyr Pro Leu Ile Phe Tyr Arg Asn Hi - #s Thr Leu Arg Gly Val                       475      - #           480      - #           485                  - - TTC GGG ACA ATG CTT GAT GGT GAA CAA GCA AG - #A CTT AAC CCT GCG TCT         1600                                                                       Phe Gly Thr Met Leu Asp Gly Glu Gln Ala Ar - #g Leu Asn Pro Ala Ser                   490          - #       495          - #       500                      - - GCA GTA TTC GAT AGC ACA TCC CGC AGT CGC AT - #A ACT CGA GTG AGT TCA         1648                                                                       Ala Val Phe Asp Ser Thr Ser Arg Ser Arg Il - #e Thr Arg Val Ser Ser               505              - #   510              - #   515                          - - AGC AGC ATC AAA GCA GCA TAC ACA ACA TCA AC - #T TGT TTT AAA GTG GTC         1696                                                                       Ser Ser Ile Lys Ala Ala Tyr Thr Thr Ser Th - #r Cys Phe Lys Val Val           520                 5 - #25                 5 - #30                 5 -      #35                                                                              - - AAG ACC AAT AAG ACC TAT TGT CTC AGC ATT GC - #T GAA ATA TCT AAT        ACT     1744                                                                    Lys Thr Asn Lys Thr Tyr Cys Leu Ser Ile Al - #a Glu Ile Ser Asn Thr                          540  - #               545  - #               550              - - CTC TTC GGA GAA TTC AGA ATC GTC CCG TTA CT - #A GTT GAG ATC CTC AAA         1792                                                                       Leu Phe Gly Glu Phe Arg Ile Val Pro Leu Le - #u Val Glu Ile Leu Lys                       555      - #           560      - #           565                  - - GAT GAC GGG GTT AGA GAA GCC AGG TCT GGC TA - #GTTGAGTC AACTATGAAA           1842                                                                       Asp Asp Gly Val Arg Glu Ala Arg Ser Gly                                               570          - #       575                                             - - GAGTTGGAAA GATGGCATTG TATCACCTAT CTTCTGCGAC ATCAAGAATC AA -             #ACCGAATG   1902                                                                 - - CCGGC                 - #                  - #                  -      #          1907                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:30:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 577 amino - #acids                                                (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                              - - Met Asp Arg Ala Val Ser Gln Val Ala Leu Gl - #u Asn Asp Glu Arg        Glu                                                                               1               5 - #                 10 - #                 15             - - Ala Lys Asn Thr Trp Arg Leu Ile Phe Arg Il - #e Ala Ile Leu Phe Leu                   20     - #             25     - #             30                  - - Thr Val Val Thr Leu Ala Ile Ser Val Ala Se - #r Leu Leu Tyr Ser Met               35         - #         40         - #         45                      - - Gly Ala Ser Thr Pro Ser Asp Leu Val Gly Il - #e Pro Thr Arg Ile Ser           50             - #     55             - #     60                          - - Arg Ala Glu Glu Lys Ile Thr Ser Thr Leu Gl - #y Ser Asn Gln Asp Val       65                 - # 70                 - # 75                 - # 80       - - Val Asp Arg Ile Tyr Lys Gln Val Ala Leu Gl - #u Ser Pro Leu Ala Leu                       85 - #                 90 - #                 95              - - Leu Asn Thr Glu Thr Thr Ile Met Asn Ala Il - #e Thr Ser Leu Ser Tyr                  100      - #           105      - #           110                  - - Gln Ile Asn Gly Ala Ala Asn Asn Ser Gly Tr - #p Gly Ala Pro Ile His              115          - #       120          - #       125                      - - Asp Pro Asp Tyr Ile Gly Gly Ile Gly Lys Gl - #u Leu Ile Val Asp Asp          130              - #   135              - #   140                          - - Ala Ser Asp Val Thr Ser Phe Tyr Pro Ser Al - #a Phe Gln Glu His Leu      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Asn Phe Ile Pro Ala Pro Thr Thr Gly Ser Gl - #y Cys Thr Arg Ile        Pro                                                                                             165  - #               170  - #               175             - - Ser Phe Asp Met Ser Ala Thr His Tyr Cys Ty - #r Thr His Asn Val Ile                  180      - #           185      - #           190                  - - Leu Ser Gly Cys Arg Asp His Ser His Ser Hi - #s Gln Tyr Leu Ala Leu              195          - #       200          - #       205                      - - Gly Val Leu Arg Thr Ser Ala Thr Gly Arg Va - #l Phe Phe Ser Thr Leu          210              - #   215              - #   220                          - - Arg Ser Ile Asn Leu Asp Asp Thr Gln Asn Ar - #g Lys Ser Cys Ser Val      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Ser Ala Thr Pro Leu Gly Cys Asp Met Leu Cy - #s Ser Lys Ala Thr        Glu                                                                                             245  - #               250  - #               255             - - Thr Glu Glu Glu Asp Tyr Asn Ser Ala Val Pr - #o Thr Arg Met Val His                  260      - #           265      - #           270                  - - Gly Arg Leu Gly Phe Asp Gly Gln Tyr His Gl - #u Lys Asp Leu Asp Val              275          - #       280          - #       285                      - - Thr Thr Leu Phe Gly Asp Trp Val Ala Asn Ty - #r Pro Gly Val Gly Gly          290              - #   295              - #   300                          - - Gly Ser Phe Ile Asp Ser Arg Val Trp Phe Se - #r Val Tyr Gly Gly Leu      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Lys Pro Asn Thr Pro Ser Asp Thr Val Gln Gl - #u Gly Lys Tyr Val        Ile                                                                                             325  - #               330  - #               335             - - Tyr Lys Arg Tyr Asn Asp Thr Cys Pro Asp Gl - #u Gln Asp Tyr Gln Ile                  340      - #           345      - #           350                  - - Arg Met Ala Lys Ser Ser Tyr Lys Pro Gly Ar - #g Phe Gly Gly Lys Arg              355          - #       360          - #       365                      - - Ile Gln Gln Ala Ile Leu Ser Ile Lys Val Se - #r Thr Ser Leu Gly Glu          370              - #   375              - #   380                          - - Asp Pro Val Leu Thr Val Pro Pro Asn Thr Va - #l Thr Leu Met Gly Ala      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Glu Gly Arg Ile Leu Thr Val Gly Thr Ser Hi - #s Phe Leu Tyr Gln        Arg                                                                                             405  - #               410  - #               415             - - Gly Ser Ser Tyr Phe Ser Pro Ala Leu Leu Ty - #r Pro Met Thr Val Ser                  420      - #           425      - #           430                  - - Asn Lys Thr Ala Thr Leu His Ser Pro Tyr Th - #r Phe Asn Ala Phe Thr              435          - #       440          - #       445                      - - Arg Pro Gly Ser Ile Pro Cys Gln Ala Ser Al - #a Arg Cys Pro Asn Ser          450              - #   455              - #   460                          - - Cys Val Thr Gly Val Tyr Thr Asp Pro Tyr Pr - #o Leu Ile Phe Tyr Arg      465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - Asn His Thr Leu Arg Gly Val Phe Gly Thr Me - #t Leu Asp Gly Glu        Gln                                                                                             485  - #               490  - #               495             - - Ala Arg Leu Asn Pro Ala Ser Ala Val Phe As - #p Ser Thr Ser Arg Ser                  500      - #           505      - #           510                  - - Arg Ile Thr Arg Val Ser Ser Ser Ser Ile Ly - #s Ala Ala Tyr Thr Thr               515         - #        520         - #        525                     - - Ser Thr Cys Phe Lys Val Val Lys Thr Asn Ly - #s Thr Tyr Cys Leu Ser          530              - #   535              - #   540                          - - Ile Ala Glu Ile Ser Asn Thr Leu Phe Gly Gl - #u Phe Arg Ile Val Pro      545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - Leu Leu Val Glu Ile Leu Lys Asp Asp Gly Va - #l Arg Glu Ala Arg        Ser                                                                                             565  - #               570  - #               575             - - Gly                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:31:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 57 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: circular                                               - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Plasmid                                                - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 538-46.26 (J - #unction A)                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                              - - CACATACGAT TTAGGTGACA CTATAGAATA CAAGCTTTAT ACCATTATAG AT - #ACATT            57                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:32:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 108 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: circular                                               - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Plasmid                                                - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 538-46.26 (J - #unction B)                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: exon                                                            (B) LOCATION: 88..102                                                         (D) OTHER INFORMATION: - #/codon.sub.-- start= 88                                  /function=- # "Translational start of hybrid protein"                         /product=- # "N-terminal peptide"                                             /number= - #1                                                                 /standard.sub.-- - #name= "Translation of synthetic DNA                       sequence"                                                       - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 103..108                                                        (C) IDENTIFICATION METHOD: - # experimental                                   (D) OTHER INFORMATION: - #/partial                                                 /codon.sub.-- - #start= 103                                                   /product=- # "NDV Heamagglutinin-Neuraminidase"                               /evidence=- # EXPERIMENTAL                                                    /gene= - #"HN"                                                                /number= - #2                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                              - - CATGTAGTCG ACTCTAGAAA AAATTGAAAA ACTATTCTAA TTTATTGCAC GG -             #AGATCTTT     60                                                                 - - TTTTTTTTTT TTTTTTTTGG CATATAAATG AATTCGGATC CG GAC CGC - #                   108                                                                                        - #                  - #           Asp Arg                                    - #                  - #             1                       - -  - - (2) INFORMATION FOR SEQ ID NO:33:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                              - - Asp Arg                                                                    1                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:34:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 108 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: circular                                               - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Plasmid                                                - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 538-46.26 (J - #unction C)                                - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 70..84                                                          (D) OTHER INFORMATION: - #/codon.sub.-- start= 70                                  /function=- # "Translational start of hybrid protein"                         /product=- # "N-terminal peptide"                                             /number= - #1                                                                 /standard.sub.-- - #name= "Translation of synthetic DNA                       sequence"                                                       - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 85..108                                                         (C) IDENTIFICATION METHOD: - # experimental                                   (D) OTHER INFORMATION: - #/partial                                                 /codon.sub.-- - #start= 85                                                    /function=- # "marker enzyme"                                                 /product=- # "Beta-galactosidase"                                             /evidence=- # EXPERIMENTAL                                                    /gene= - #"lacZ"                                                              /number= - #2                                                                 /citation=- # ([1])                                             - -      (x) PUBLICATION INFORMATION:                                                  (A) AUTHORS: Ferrari, F - #ranco A                                                 Trach, Ka - #thleen                                                           Hoch, Jam - #es A                                                        (B) TITLE: Sequence Ana - #lysis of the spo0B Locus Reveals                        a Polycis - #tronic Transcription Unit                                   (C) JOURNAL: J. Bacteri - #ol.                                                (D) VOLUME: 161                                                               (E) ISSUE: 2                                                                  (F) PAGES: 556-562                                                            (G) DATE: Feb.-1985                                                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                              - - TGCGACATCA AGAATCAAAC CGAATGCCCT CGACTCTAGA ATTTCATTTT GT -             #TTTTTTCT     60                                                                 - - ATGCTATAA ATG AAT TCG GAT CCC GTC GTT TTA CAA - # CGT CGT GAC TGG            108                                                                                Met Asn Ser Asp Pro - #Val Val Leu Gln Arg Arg Asp Trp                          1      - #         5   1      - #         5                        - -  - - (2) INFORMATION FOR SEQ ID NO:35:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 5 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                              - - Met Asn Ser Asp Pro                                                        1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:36:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 8 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:                              - - Val Val Leu Gln Arg Arg Asp Trp                                             1               - #5                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:37:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 108 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: circular                                               - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Plasmid                                                - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 538-46.26                                                 - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..54                                                           (C) IDENTIFICATION METHOD: - # experimental                                   (D) OTHER INFORMATION: - #/partial                                                 /codon.sub.-- - #start= 1                                                     /function=- # "marker enzyme"                                                 /product=- # "Beta-galactosidase"                                             /evidence=- # EXPERIMENTAL                                                    /gene= - #"lacZ"                                                              /number= - #1                                                                 /citation=- # ([1])                                             - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 55..63                                                          (C) IDENTIFICATION METHOD: - # experimental                                   (D) OTHER INFORMATION: - #/codon.sub.-- start= 55                                  /function=- # "Translational finish of hybrid                                 protein"                                                                      /product=- # "C-terminal peptide"                                             /evidence=- # EXPERIMENTAL                                                    /number= - #2                                                                 /standard.sub.-- - #name= "Translation of synthetic DNA                       sequence"                                                       - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:                              - - GAA ATC CAG CTG AGC GCC GGT CGC TAC CAT TA - #C CAG TTG GTC TGG TGT           48                                                                       Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys             1               5 - #                 10 - #                 15              - - CAA AAA GAT CCA TAATTAATTA ACCCGGGTCG AGGGTCGAAG AC - #CAAATTCT              100                                                                       Gln Lys Asp Pro                                                                         1                                                                    - - AACATGGT                - #                  - #                  -     #         108                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:38:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 18 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:                              - - Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys        1               5 - #                 10 - #                 15              - - Gln Lys                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:39:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 2 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:                              - - Asp Pro                                                                    1                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:40:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 57 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: circular                                               - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Plasmid                                                - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 538-46.26 (J - #unction E)                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:                              - - AGATCCCCGG GCGAGCTCGA ATTCGTAATC ATGGTCATAG CTGTTTCCTG TG - #TGAAA            57                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:41:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 27 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: N                                                  - -     (iv) ANTI-SENSE: N                                                    - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Pseudorabies - # virus \ Synthetic oligonucleotid                   primer                                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:                              - - CGCGAATTCG CTCGCAGCGC TATTGGC          - #                  - #                 27                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:42:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 19 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: N                                                  - -     (iv) ANTI-SENSE: N                                                    - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Pseudorabies - # virus \ Synthetic oligonucleotid    e                                                                                            primer                                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:                              - - GTAGGAGTGG CTGCTGAAG             - #                  - #                      - # 19                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:43:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 70 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:                              - - AAAAATTGAA AAACTATTCT AATTTATTGC ACGGAGATCT TTTTTTTTTT TT -             #TTTTTTTG     60                                                                 - - GCATATAAAT                - #                  - #                      - #        70                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:44:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 74 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:                              - - TTTTTTTTTT TTTTTTTTTT GGCATATAAA TAGATCTGTA TCCTAAAATT GA -             #ATTGTAAT     60                                                                 - - TATCGATAAT AAAT              - #                  - #                      - #     74                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:45:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 37 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:                              - - GTATCCTAAA ATTGAATTGT AATTATCGAT AATAAAT      - #                       - #      37                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:46:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 41 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:                              - - CGACTCTAGA ATTTCATTTT GTTTTTTTCT ATGCTATAAA T    - #                      - #   41                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:47:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 60 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:                              - - CACATACGAT TTAGGTGACA CTATAGAATA CAAGCTTTGA GTCTATTGGT TA -             #TTTATACG     60                                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:48:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 123 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 100..123                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:                              - - TGAATATATA GCAAATAAAG GAAAAATTGT TATCGTTGCT GCATTAGATG GA -            #ACATAGGT     60                                                                 - - CGACTCTAGA ATTTCATTTT GTTTTTTTCT ATGCTATAA ATG AAT TCG - # GAT CCC           114                                                                                        - #                  - #       Met Asn Ser Asp Pro                            - #                  - #         1         - #      5        - - GTC GTT TTA              - #                  - #                       - #        123                                                                  Val Val Leu                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:49:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 8 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:                              - - Met Asn Ser Asp Pro Val Val Leu                                            1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:50:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 132 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..63                                                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:                              - - GAA ATC CAG CTG AGC GCC GGT CGC TAC CAT TA - #C CAG TTG GTC TGG        TGT       48                                                                    Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys            1               5 - #                 10 - #                 15              - - CAA AAA GAT CCA TAATTAATTA ACCCGGGTCG ACCTATGAAC GT - #AAACCATT              100                                                                       Gln Lys Asp Pro                                                                            20                                                                - - TGGTAATATT CTTAATCTTA TACCATTATC GG       - #                  - #             132                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:51:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:                              - - Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys        1               5 - #                 10 - #                 15              - - Gln Lys Asp Pro                                                                       20                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:52:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 66 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:                              - - TCTACTATTG TATATATAGG ATCCCCGGGC GAGCTCGAAT TCGTAATCAT GG -             #TCATAGCT     60                                                                 - - GTTTCC                 - #                  - #                  -     #           66                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:53:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:                              - - ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:54:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 104 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 81..104                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:                              - - AAATATATAA ATACCATGTT AGAATTTGGT CTGCTGCAGG TCGACTCTAG AA -             #TTTCATTT     60                                                                 - - TGTTTTTTTC TATGCTATAA ATG AAT TCG GAT CCC GTC GT - #T TTA                    - #104                                                                                     - #    Met Asn Ser Asp Pro Val Val Leu                                        - #      1            - #   5                                - -  - - (2) INFORMATION FOR SEQ ID NO:55:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 8 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:                              - - Met Asn Ser Asp Pro Val Val Leu                                            1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:56:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 150 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..63                                                  - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 130..150                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:                              - - GAA ATC CAG CTG AGC GCC GGT CGC TAC CAT TA - #C CAG TTG GTC TGG TGT           48                                                                       Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys             1               5 - #                 10 - #                 15              - - CAA AAA GAT CCA TAATTAATTA ACCCGGTCGA CTCTAGAAAG AT - #CTGTATCC              100                                                                       Gln Lys Asp Pro                                                                            20                                                                - - TAAAATTGAA TTGTAATTAT CGATAATAA ATG AAT TCC GGC ATG - # GCC TCG              150                                                                                         - #              Met Asn S - #er Gly Met Ala Ser                              - #                1  - #             5                      - -  - - (2) INFORMATION FOR SEQ ID NO:57:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57:                              - - Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys        1               5 - #                 10 - #                 15              - - Gln Lys Asp Pro                                                                       20                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:58:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:                              - - Met Asn Ser Gly Met Ala Ser                                                1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:59:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 109 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:                              - - CCATGCTCTA GAGGATCCCC GGGCGAGCTC GAATTCGGAT CCATAATTAA TT -             #AATTAATT     60                                                                 - - TTTATCCCGG GTCGACCGGG TCGACCTGCA GCCTACATGG AAATCTACC  - #                  109                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:60:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60:                              - - TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:61:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61:                              - - ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:62:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 104 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 81..104                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62:                              - - AAATATATAA ATACCATGTT AGAATTTGGT CTGCTGCAGG TCGACTCTAG AA -             #TTTCATTT     60                                                                 - - TGTTTTTTTC TATGCTATAA ATG AAT TCG GAT CCC GTC GT - #T TTA                    - #104                                                                                     - #    Met Asn Ser Asp Pro Val Val Leu                                        - #      1            - #   5                                - -  - - (2) INFORMATION FOR SEQ ID NO:63:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 8 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:63:                              - - Met Asn Ser Asp Pro Val Val Leu                                            1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:64:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 182 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..63                                                  - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 156..182                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64:                              - - GAA ATC CAG CTG AGC GCC GGT CGC TAC CAT TA - #C CAG TTG GTC TGG TGT           48                                                                       Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys             1               5 - #                 10 - #                 15              - - CAA AAA GAT CCA TAATTAATTA ACCCGGTCGA CTCTAGAAAA AA - #TTGAAAAA              100                                                                       Gln Lys Asp Pro                                                                            20                                                                - - CTATTCTAAT TTATTGCACG GAGATCTTTT TTTTTTTTTT TTTTTTGGCA TA - #TAA        ATG     158                                                                                       - #                  - #                  - #             Met                                                                                               - #                  - #                  - #             1                                                                               - - AAT TCC GGC ATG GCC TCG CTC GCG     - #                  - #                   182                                                                     Asn Ser Gly Met Ala Ser Leu Ala                                                             5                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:65:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:65:                              - - Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys        1               5 - #                 10 - #                 15              - - Gln Lys Asp Pro                                                                       20                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:66:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 9 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:                              - - Met Asn Ser Gly Met Ala Ser Leu Ala                                        1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:67:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 109 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:67:                              - - CCATGCTCTA GAGGATCCCC GGGCGAGCTC GAATTCGGAT CCATAATTAA TT -             #AATTAATT     60                                                                 - - TTTATCCCGG GTCGACCGGG TCGACCTGCA GCCTACATGG AAATCTACC  - #                  109                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:68:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68:                              - - TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:69:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:69:                              - - ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:70:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 104 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 81..104                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:70:                              - - AAATATATAA ATACCATGTT AGAATTTGGT CTGCTGCAGG TCGACTCTAG AA -             #TTTCATTT     60                                                                 - - TGTTTTTTTC TATGCTATAA ATG AAT TCG GAT CCC GTC GT - #T TTA                    - #104                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:71:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 8 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:71:                              - - Met Asn Ser Asp Pro Val Val Leu                                            1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:72:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 180 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..63                                                  - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 160..180                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:72:                              - - GAA ATC CAG CTG AGC GCC GGT CGC TAC CAT TA - #C CAG TTG GTC TGG TGT           48                                                                       Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys             1               5 - #                 10 - #                 15              - - CAA AAA GAT CCA TAATTAATTA ACCCGGTCGA CTCTAGATTT TT - #TTTTTTTT              100                                                                       Gln Lys Asp Pro                                                                            20                                                                - - TTTTTTTGGC ATATAAATAG ATCTGTATCC TAAAATTGAA TTGTAATTAT CG -             #ATAATAA     159                                                                 - - ATG AAT TCC GGC ATG GCC TCG       - #                  - #                     180                                                                    Met Asn Ser Gly Met Ala Ser                                                     1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:73:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:73:                              - - Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys        1               5 - #                 10 - #                 15              - - Gln Lys Asp Pro                                                                       20                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:74:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:74:                              - - Met Asn Ser Gly Met Ala Ser                                                1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:75:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 109 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:75:                              - - CCATGCTCTA GAGGATCCCC GGGCGAGCTC GAATTCGGAT CCATAATTAA TT -             #AATTAATT     60                                                                 - - TTTATCCCGG GTCGACCGGG TCGACCTGCA GCCTACATGG AAATCTACC  - #                  109                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:76:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:76:                              - - TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:77:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:77:                              - - ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:78:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 117 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 94..117                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:78:                              - - GGTCTGCTGC AGGTCGACTC TAGAAAAAAT TGAAAAACTA TTCTAATTTA TT -             #GCACGGAG     60                                                                 - - ATCTTTTTTT TTTTTTTTTT TTTTGGCATA TAA ATG AAT TCC GG - #C TTC AGT       AAC ATA 117                                                                                       - #                  - # Met Asn Ser Gly Phe Ser Asn      Ile                                                                                               - #                  - #  1               5 - #              8                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:79:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 8 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:79:                              - - Met Asn Ser Gly Phe Ser Asn Ile                                            1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:80:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 126 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 103..126                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:80:                              - - CGCAACATAC CTAACTGCTT CATTTCTGAT CCATAATTAA TTAATTTTTA TC -             #CCGGCGCG     60                                                                 - - CCTCGACTCT AGAATTTCAT TTTGTTTTTT TCTATGCTAT AA ATG AAT - # TCG GAT           114                                                                                        - #                  - #            Met Asn Ser As -       #p                                                                                                - #                  - #             1                       - - CCC GTC GTT TTA            - #                  - #                      - #      126                                                                 Pro Val Val Leu                                                                 5                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:81:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 8 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:81:                              - - Met Asn Ser Asp Pro Val Val Leu                                            1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:82:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 96 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..63                                                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:82:                              - - GAA ATC CAG CTG AGC GCC GGT CGC TAC CAT TA - #C CAG TTG GTC TGG TGT           48                                                                       Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys             1               5 - #                 10 - #                 15              - - CAA AAA GAT CCA TAATTAATTA ACCCGGGTCG ACCTGCAGCC TA - #CATG                   96                                                                       Gln Lys Asp Pro                                                                            20                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:83:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:83:                              - - Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys        1               5 - #                 10 - #                 15              - - Gln Lys Asp Pro                                                                       20                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:84:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:84:                              - - TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:85:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:85:                              - - ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:86:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 124 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 104..124                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:86:                              - - GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TG -             #GCATATAA     60                                                                 - - ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAA ATG AA - #T TCG        CTA CTT  118                                                                                      - #                  - #            Met Asn Ser Le -      #u Leu                                                                                            - #                  - #              1    - #               5                                                                            - - GGA ACT                - #                  - #                  -      #          124                                                                  Gly Thr                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:87:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:87:                              - - Met Asn Ser Leu Leu Gly Thr                                                1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:88:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 126 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..12                                                  - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 103..126                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:88:                              - - ATA AAA ATG TGATTAAGTC TGAATGTGGA TCCATAATTA ATTAATTTT - #T                   49                                                                      Ile Lys Met                                                                     1                                                                            - - ATCCCGGCGC GCCTCGACTC TAGAATTTCA TTTTGTTTTT TTCTATGCTA TA - #A ATG           105                                                                                         - #                  - #                  - #     Met                         - #                  - #                  - #       1        - - AAT TCG GAT CCC GTC GTT TTA       - #                  - #                     126                                                                     Asn Ser Asp Pro Val Val Leu                                                                 5                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:89:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 3 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:89:                              - - Ile Lys Met                                                                1                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:90:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 8 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:90:                              - - Met Asn Ser Asp Pro Val Val Leu                                            1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:91:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 116 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..63                                                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:91:                              - - GAA ATC CAG CTG AGC GCC GGT CGC TAC CAT TA - #C CAG TTG GTC TGG TGT           48                                                                       Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys             1               5 - #                 10 - #                 15              - - CAA AAA GAT CCA TAATTAATTA ACCCGGGTCG AGGCGCGCCG GG - #TCGACCTG              100                                                                       Gln Lys Asp Pro                                                                            20                                                                - - CAGGCGGCCG CTATAC             - #                  - #                      - #   116                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:92:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:92:                              - - Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys        1               5 - #                 10 - #                 15              - - Gln Lys Asp Pro                                                                       20                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:93:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:93:                              - - TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:94:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:94:                              - - ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:95:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 124 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 104..124                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:95:                              - - GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TG -             #GCATATAA     60                                                                 - - ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAA ATG AA - #T TCC        CCT GCC  118                                                                                      - #                  - #            Met Asn Ser Pr -      #o Ala                                                                                            - #                  - #              1    - #               5                                                                            - - GCC CGG                - #                  - #                  -      #          124                                                                  Ala Arg                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:96:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 7 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:96:                              - - Met Asn Ser Pro Ala Ala Arg                                                1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:97:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 126 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..36                                                  - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 103..126                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:97:                              - - CTC CAG GAG CCC GCT CGC CTC GAG CGG GAT CC - #A TAATTAATTA             ATTTTTATCC     53                                                               Leu Gln Glu Pro Ala Arg Leu Glu Arg Asp Pr - #o                                 1               5 - #                 10                                     - - CGGCGCGCCT CGACTCTAGA ATTTCATTTT GTTTTTTTCT ATGCTATAA ATG - # AAT            108                                                                                        - #                  - #                  - #Met Asn                          - #                  - #                  - #  1             - - TCG GAT CCC GTC GTT TTA         - #                  - #                      - # 126                                                                  Ser Asp Pro Val Val Leu                                                                 5                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:98:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 11 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:98:                              - - Leu Gln Glu Pro Ala Arg Leu Glu Arg Asp Pr - #o                            1               5 - #                 10                                     - -  - - (2) INFORMATION FOR SEQ ID NO:99:                                    - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 8 amino - #acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:99:                              - - Met Asn Ser Asp Pro Val Val Leu                                            1               5                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:100:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 116 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..63                                                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:100:                             - - GAA ATC CAG CTG AGC GCC GGT CGC TAC CAT TA - #C CAG TTG GTC TGG TGT           48                                                                       Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys              1               - #5                  - #10                  - #15          - - CAA AAA GAT CCA TAATTAATTA ACCCGGGTCG AGGCGCGCCG GG - #TCGACCTG              100                                                                       Gln Lys Asp Pro                                                                            20                                                                - - CAGGCGGCCG CTATAC             - #                  - #                      - #   116                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:101:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino - #acids                                                 (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: protein                                           - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:101:                             - - Glu Ile Gln Leu Ser Ala Gly Arg Tyr His Ty - #r Gln Leu Val Trp Cys        1               5 - #                 10 - #                 15              - - Gln Lys Asp Pro                                                                       20                                                                - -  - - (2) INFORMATION FOR SEQ ID NO:102:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:102:                             - - TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:103:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 30 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:103:                             - - CCGAATTCCG GCTTCAGTAA CATAGGATCG         - #                  - #               30                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:104:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:104:                             - - GTACCCATAC TGGTCGTGGC            - #                  - #                      - # 20                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:105:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 26 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:105:                             - - CCGGAATTCG CTACTTGGAA CTCTGG          - #                  - #                  26                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:106:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:106:                             - - CATTGTCCCG AGACGGACAG            - #                  - #                      - # 20                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:107:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 19 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:107:                             - - CGCGATCCAA CTATCGGTG             - #                  - #                      - # 19                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:108:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 26 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:108:                             - - GCGGATCCAC ATTCAGACTT AATCAC          - #                  - #                  26                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:109:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 30 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:109:                             - - ATGAATTCCC CTGCCGCCCG GACCGGCACC         - #                  - #               30                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:110:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 30 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:110:                             - - CATGGATCCC GCTCGAGGCG AGCGGGCTCC         - #                  - #               30                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:111:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 42 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:111:                             - - CTGGTTCGGC CCAGAATTCT ATGGGTCTCG CGCGGCTCGT GG    - #                      - #  42                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:112:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 42 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:112:                             - - CTCGCTCGCC CAGGATCCCT AGCGGAGGAT GGACTTGAGT CG    - #                      - #  42                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:113:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 3628 base - #pairs                                                (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 57..1226                                               - -     (ix) FEATURE:                                                                  (A) NAME/KEY: CDS                                                             (B) LOCATION: 1362..3395                                             - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:113:                             - - TTGAAGATGA ATGCATAGAG GAAGATGATG TCGANACGTC ATTATTTAAT GT -             #ATAAATGG     60                                                                 - - ATAAATTGTA TGCGGCAATA TTCGGCGTTT TTATGACATC TAAAGATGAT GA -            #TTTTAATA    120                                                                 - - ACTTTATAGA AGTTGTAAAA TCTGTATTAA CAGATACATC ANCTAATCAT AC -            #AATATCGT    180                                                                 - - CGTCCAATAA TAATACATGG ATATATATAT TTCTAGCGAT ATTATTTGGT GT -            #TATGGNAT    240                                                                 - - TATTAGTTTT TANTTTGTAT GTAGAAGTTC CTAAACCNAC TTANATGGAG GA -            #AGCAGATA    300                                                                 - - ACCNACTCGT TNTAAATAGT ATTAGTGCTA GAGCATTGGN GGCATTTTTT GT -            #ATCTAAAA    360                                                                 - - NTANTGATAT GGTCGNTGAA NTAGTTNCCC AAAAATNTCC NCCAAAGAAG AN -            #ATCACAAA    420                                                                 - - TAAAACGCAT AGATACACGA ATTCCTATTG ATCTTATTAA TCAACAATTC GT -            #TAAAAGAT    480                                                                 - - TTAAACTAGA AAATTATAAA AATGGAATTT TATCCGTTCT TATCAATAGT TT -            #AGTCGAAA    540                                                                 - - ATAATTACTT TGAACAAGAT GGTAAACTTA ATAGCAGTGA TATTGATGAA TT -            #AGTGCTCA    600                                                                 - - CAGACATAGA GAAAAAGATT TTATCGTTGA TTCCTAGATG TTCTCCTCTT TA -            #TATAGATA    660                                                                 - - TCAGTGACGT TAAAGTTCTC GCATCTAGGT TAANNAAAAG TGCTAAATCA TT -            #TACGTTTA    720                                                                 - - ATGATCATGA ATATATTATA CAATCTGATA AAATAGAGGA ATTAATAAAT AG -            #TTTATCTA    780                                                                 - - GAAACCATGA TATTATACTA GATGAAAAAA GTTCTATTAA AGACAGCATA TA -            #TATACTAT    840                                                                 - - CTGATGATCT TTTGAATATA CTTCGTGAAA GATTATTTAG ATGTCCACAG GT -            #TAAAGATA    900                                                                 - - ATACTATTTC TAGAACACGT CTATATGATT ATTTTACTAG AGTGTCAAAG AA -            #AGAAGAAG    960                                                                 - - CGAAAATATA CGTTATATTG AAAGATTTAA AGATTGCTGA TATACTCGGT AT -            #CGAAACAG   1020                                                                 - - TAACGATAGG ATCATTTGTA TATACGAAAT ATAGCATGTT GATTAATTCA AT -            #TTCGTCTA   1080                                                                 - - ATGTTGATAG ATATTCAAAA AGGTTCCATG ACTCTTTTTA TGAAGATATT GC -            #GGAATTTA   1140                                                                 - - TAAAGGATAA TGAAAAAATT AATGTATCCA GAGTTGTTGA ATGCCTTATC GT -            #ACCTAATA   1200                                                                 - - TTAATATAGA GTTATTAACT GAATAAGTAT ATATAAATGA TTGTTTTTAT AA -            #TGTTTGTT   1260                                                                 - - ATCGCATTTA GTTTTGCTGT ATGGTTATCA TATACATTTT TAAGGCCGTA TA -            #TGATAAAT   1320                                                                 - - GAAAATATAT AAGCACTTAT TTTTGTTAGT ATAATAACAC AATGCCGTCG TA -            #TATGTATC   1380                                                                 - - CGAAGAACGC AAGAAAAGTA ATTTCAAAGA TTATATCATT ACAACTTGAT AT -            #TAAAAAAC   1440                                                                 - - TTCCTAAAAA ATATATAAAT ACCATGTTAG AATTTGGTCT ACATGGAAAT CT -            #ACCAGCTT   1500                                                                 - - GTATGTATAA AGATGCCGTA TCATATGATA TAAATAATAT AAGATTTTTA CC -            #TTATAATT   1560                                                                 - - GTGTTATGGT TAAAGATTTA ATAAATGTTA TAAAATCATC ATCTGTAATA GA -            #TACTAGAT   1620                                                                 - - TACATCAATC TGTATTAAAA CATCGTAGAG CGTTAATAGA TTACGGCGAT CA -            #AGACATTA   1680                                                                 - - TCACTTTAAT GATCATTAAT AAGTTACTAT CGATAGATGA TATATCCTAT AT -            #ATTAGATA   1740                                                                 - - AAAAAATAAT TCATGTAACA AAAATATTAA AAATAGACCC TACAGTAGCC AA -            #TTCAAACA   1800                                                                 - - TGAAACTGAA TAAGATAGAG CTTGTAGATG TAATAACATC AATACCTAAG TC -            #TTCCTATA   1860                                                                 - - CATATTTATA TAATAATATG ATCATTGATC TCGATACATT ATTATATTTA TC -            #CGATGCAT   1920                                                                 - - TCCACATACC CCCCACACAT ATATCATTAC GTTCACTTAG AGATATAAAC AG -            #GATTATTG   1980                                                                 - - AATTGCTTAA AAAATATCCG AATAATAATA TTATTGATTA TATATCCGAT AG -            #CATAAAAT   2040                                                                 - - CAAATAGTTC ATTCATTCAC ATACTTCATA TGATAATATC AAATATGTTT CC -            #TGCTATAA   2100                                                                 - - TCCCTAGTGT AAACGATTTT ATATCTACCG TAGTTGATAA AGATCGACTT AT -            #TAATATGT   2160                                                                 - - ATGGGATTAA GTGTGTTGCT ATGTTTTCGT ACGATATAAA CATGATCGAT TT -            #AGAGTCAT   2220                                                                 - - TAGATGACTC AGATTACATA TTTATAGAAA AAAATATATC TATATACGAC GT -            #TAAATGTA   2280                                                                 - - GAGATTTTGC GAATATGATT AGAGATAAGG TTAAAAGAGA AAAGAATAGA AT -            #ATTAACTA   2340                                                                 - - CGAAATGTGA AGATATTATA AGATATATAA AATTATTCAG TAAAAATAGA AT -            #AAACGATG   2400                                                                 - - AAAATAATAA GGTGGAGGAG GTGTTGATAC ATATTGATAA TGTATCTAAA AA -            #TAATAAAT   2460                                                                 - - TATCACTGTC TGATATATCA TCTTTAATGG ATCAATTTCG TTTAAATCCA TG -            #TACCATAA   2520                                                                 - - GAAATATATT ATTATCTTCA GCAACTATAA AATCAAAACT ATTAGCGTTA CG -            #GGCAGTAA   2580                                                                 - - AAAACTGGAA ATGTTATTCA TTGACAAATG TATCAATGTA TAAAAAAATA AA -            #GGGTGTTA   2640                                                                 - - TCGTAATGGA TATGGTTGAT TATATATCTA CTAACATTCT TAAATACCAT AA -            #ACAATTAT   2700                                                                 - - ATGATAAAAT GAGTACGTTT GAATATAAAC GAGATATTAA ATCATGTAAA TG -            #CTCGATAT   2760                                                                 - - GTTCCGACTC TATAACACAT CATATATATG AAACAACATC ATGTATAAAT TA -            #TAAATCTA   2820                                                                 - - CCGATAATGA TCTTATGATA GTATTGTTCA ATCTAACTAG ATATTTAATG CA -            #TGGGATGA   2880                                                                 - - TACATCCTAA TCTTATAAGC GTAAAAGGAT GGGGTCCCCT TATTGGATTA TT -            #AACGGGTG   2940                                                                 - - ATATAGGTAT TAATTTAAAA CTATATTCCA CCATGAATAT AAATGGGCTA CG -            #GTATGGAG   3000                                                                 - - ATATTACGTT ATCTTCATAC GATATGAGTA ATAAATTAGT CTCTATTATT AA -            #TACACCCA   3060                                                                 - - TATATGAGTT AATACCGTTT ACTACATGTT GTTCACTCAA TGAATATTAT TC -            #AAAAATTG   3120                                                                 - - TGATTTTAAT AAATGTTATT TTAGAATATA TGATATCTAT TATATTATAT AG -            #AATATTGA   3180                                                                 - - TCGTAAAAAG ATTTAATAAC ATTAAAGAAT TTATTTCAAA AGTCGTAAAT AC -            #TGTACTAG   3240                                                                 - - AATCATCAGG CATATATTTT TGTCAGATGC GTGTACATGA ACAAATTGAA TT -            #GGAAATAG   3300                                                                 - - ATGAGCTCAT TATTAATGGA TCTATGCCTG TACAGCTTAT GCATTTACTT CT -            #AAAGGTAG   3360                                                                 - - CTACCATAAT ATTAGAGGAA ATCAAAGAAA TATAACGTAT TTTTTCTTTT AA -            #ATAAATAA   3420                                                                 - - AAATACTTTT TTTTTTAAAC AAGGGGTGCT ACCTTGTCTA ATTGTATCTT GT -            #ATTTTGGA   3480                                                                 - - TCTGATGCAA GATTATTAAA TAATCGTATG AAAAAGTAGT AGATATAGTT TA -            #TATCGTTA   3540                                                                 - - CTGGACATGA TATTATGTTT AGTTAATTCT TCTTTGGCAT GAATTCTACA CG -            #TCGGANAA   3600                                                                 - - GGTAATGTAT CTATAATGGT ATAAAGCT         - #                  - #               3628                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:114:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 389 amino - #acids                                                (B) TYPE: amino acid                                                          (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:114:                             - - Met Asp Lys Leu Tyr Ala Ala Ile Phe Gly Va - #l Phe Met Thr Ser Lys      1               5   - #                10  - #                15               - - Asp Asp Asp Phe Asn Asn Phe Ile Glu Val Va - #l Lys Ser Val Leu Thr                  20      - #            25      - #            30                   - - Asp Thr Ser Xaa Asn His Thr Ile Ser Ser Se - #r Asn Asn Asn Thr Trp              35          - #        40          - #        45                       - - Ile Tyr Ile Phe Leu Ala Ile Leu Phe Gly Va - #l Met Xaa Leu Leu Val          50              - #    55              - #    60                           - - Phe Xaa Leu Tyr Val Glu Val Pro Lys Pro Th - #r Xaa Met Glu Glu Ala      65                  - #70                  - #75                  - # 80       - - Asp Asn Xaa Leu Val Xaa Asn Ser Ile Ser Al - #a Arg Ala Leu Xaa Ala                      85  - #                90  - #                95               - - Phe Phe Val Ser Lys Xaa Xaa Asp Met Val Xa - #a Glu Xaa Val Xaa Gln                  100      - #           105      - #           110                  - - Lys Xaa Pro Pro Lys Lys Xaa Ser Gln Ile Ly - #s Arg Ile Asp Thr Arg              115          - #       120          - #       125                      - - Ile Pro Ile Asp Leu Ile Asn Gln Gln Phe Va - #l Lys Arg Phe Lys Leu          130              - #   135              - #   140                          - - Glu Asn Tyr Lys Asn Gly Ile Leu Ser Val Le - #u Ile Asn Ser Leu Val      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Glu Asn Asn Tyr Phe Glu Gln Asp Gly Lys Le - #u Asn Ser Ser Asp        Ile                                                                                             165  - #               170  - #               175             - - Asp Glu Leu Val Leu Thr Asp Ile Glu Lys Ly - #s Ile Leu Ser Leu Ile                  180      - #           185      - #           190                  - - Pro Arg Cys Ser Pro Leu Tyr Ile Asp Ile Se - #r Asp Val Lys Val Leu              195          - #       200          - #       205                      - - Ala Ser Arg Leu Xaa Lys Ser Ala Lys Ser Ph - #e Thr Phe Asn Asp His          210              - #   215              - #   220                          - - Glu Tyr Ile Ile Gln Ser Asp Lys Ile Glu Gl - #u Leu Ile Asn Ser Leu      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - -  Ser Arg Asn His Asp Ile Ile Leu Asp Glu - #Lys Ser Ser Ile Lys        Asp                                                                                             245  - #               250  - #               255             - - Ser Ile Tyr Ile Leu Ser Asp Asp Leu Leu As - #n Ile Leu Arg Glu Arg                  260      - #           265      - #           270                  - - Leu Phe Arg Cys Pro Gln Val Lys Asp Asn Th - #r Ile Ser Arg Thr Arg              275          - #       280          - #       285                      - - Leu Tyr Asp Tyr Phe Thr Arg Val Ser Lys Ly - #s Glu Glu Ala Lys Ile          290              - #   295              - #   300                          - - Tyr Val Ile Leu Lys Asp Leu Lys Ile Ala As - #p Ile Leu Gly Ile Glu      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Thr Val Thr Ile Gly Ser Phe Val Tyr Thr Ly - #s Tyr Ser Met Leu        Ile                                                                                             325  - #               330  - #               335             - - Asn Ser Ile Ser Ser Asn Val Asp Arg Tyr Se - #r Lys Arg Phe His Asp                  340      - #           345      - #           350                  - - Ser Phe Tyr Glu Asp Ile Ala Glu Phe Ile Ly - #s Asp Asn Glu Lys Ile              355          - #       360          - #       365                      - - Asn Val Ser Arg Val Val Glu Cys Leu Ile Va - #l Pro Asn Ile Asn Ile          370              - #   375              - #   380                          - - Glu Leu Leu Thr Glu                                                      385                                                                            - -  - - (2) INFORMATION FOR SEQ ID NO:115:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 677 amino - #acids                                                (B) TYPE: amino acid                                                          (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Swinepox - #virus                                               (B) STRAIN: Kasza                                                             (C) INDIVIDUAL ISOLATE: - #S-SPV-001                                 - -    (vii) IMMEDIATE SOURCE:                                                         (B) CLONE: 515-85.1                                                  - -   (viii) POSITION IN GENOME:                                                       (B) MAP POSITION:  23.2                                                       (C) UNITS: %G                                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:115:                             - - Met Pro Ser Tyr Met Tyr Pro Lys Asn Ala Ar - #g Lys Val Ile Ser Lys      1               5   - #                10  - #                15               - - Ile Ile Ser Leu Gln Leu Asp Ile Lys Lys Le - #u Pro Lys Lys Tyr Ile                  20      - #            25      - #            30                   - - Asn Thr Met Leu Glu Phe Gly Leu His Gly As - #n Leu Pro Ala Cys Met              35          - #        40          - #        45                       - - Tyr Lys Asp Ala Val Ser Tyr Asp Ile Asn As - #n Ile Arg Phe Leu Pro          50              - #    55              - #    60                           - - Tyr Asn Cys Val Met Val Lys Asp Leu Ile As - #n Val Ile Lys Ser Ser      65                  - #70                  - #75                  - #80        - -  Ser Val Ile Asp Thr Arg Leu His Gln Ser - #Val Leu Lys His Arg Arg                      85  - #                90  - #                95               - - Ala Leu Ile Asp Tyr Gly Asp Gln Asp Ile Il - #e Thr Leu Met Ile Ile                  100      - #           105      - #           110                  - - Asn Lys Leu Leu Ser Ile Asp Asp Ile Ser Ty - #r Ile Leu Asp Lys Lys              115          - #       120          - #       125                      - - Ile Ile His Val Thr Lys Ile Leu Lys Ile As - #p Pro Thr Val Ala Asn          130              - #   135              - #   140                          - - Ser Asn Met Lys Leu Asn Lys Ile Glu Leu Va - #l Asp Val Ile Thr Ser      145                 1 - #50                 1 - #55                 1 -      #60                                                                              - - Ile Pro Lys Ser Ser Tyr Thr Tyr Leu Tyr As - #n Asn Met Ile Ile        Asp                                                                                             165  - #               170  - #               175             - - Leu Asp Thr Leu Leu Tyr Leu Ser Asp Ala Ph - #e His Ile Pro Pro Thr                  180      - #           185      - #           190                  - - His Ile Ser Leu Arg Ser Leu Arg Asp Ile As - #n Arg Ile Ile Glu Leu              195          - #       200          - #       205                      - - Leu Lys Lys Tyr Pro Asn Asn Asn Ile Ile As - #p Tyr Ile Ser Asp Ser          210              - #   215              - #   220                          - - Ile Lys Ser Asn Ser Ser Phe Ile His Ile Le - #u His Met Ile Ile Ser      225                 2 - #30                 2 - #35                 2 -      #40                                                                              - - Asn Met Phe Pro Ala Ile Ile Pro Ser Val As - #n Asp Phe Ile Ser        Thr                                                                                              245 - #                250 - #                255            - - Val Val Asp Lys Asp Arg Leu Ile Asn Met Ty - #r Gly Ile Lys Cys Val                  260      - #           265      - #           270                  - - Ala Met Phe Ser Tyr Asp Ile Asn Met Ile As - #p Leu Glu Ser Leu Asp              275          - #       280          - #       285                      - - Asp Ser Asp Tyr Ile Phe Ile Glu Lys Asn Il - #e Ser Ile Tyr Asp Val          290              - #   295              - #   300                          - - Lys Cys Arg Asp Phe Ala Asn Met Ile Arg As - #p Lys Val Lys Arg Glu      305                 3 - #10                 3 - #15                 3 -      #20                                                                              - - Lys Asn Arg Ile Leu Thr Thr Lys Cys Glu As - #p Ile Ile Arg Tyr        Ile                                                                                             325  - #               330  - #               335             - - Lys Leu Phe Ser Lys Asn Arg Ile Asn Asp Gl - #u Asn Asn Lys Val Glu                  340      - #           345      - #           350                  - - Glu Val Leu Ile His Ile Asp Asn Val Ser Ly - #s Asn Asn Lys Leu Ser              355          - #       360          - #       365                      - - Leu Ser Asp Ile Ser Ser Leu Met Asp Gln Ph - #e Arg Leu Asn Pro Cys          370              - #   375              - #   380                          - - Thr Ile Arg Asn Ile Leu Leu Ser Ser Ala Th - #r Ile Lys Ser Lys Leu      385                 3 - #90                 3 - #95                 4 -      #00                                                                              - - Leu Ala Leu Arg Ala Val Lys Asn Trp Lys Cy - #s Tyr Ser Leu Thr        Asn                                                                                             405  - #               410  - #               415             - - Val Ser Met Tyr Lys Lys Ile Lys Gly Val Il - #e Val Met Asp Met Val                  420      - #           425      - #           430                  - - Asp Tyr Ile Ser Thr Asn Ile Leu Lys Tyr Hi - #s Lys Gln Leu Tyr Asp              435          - #       440          - #       445                      - - Lys Met Ser Thr Phe Glu Tyr Lys Arg Asp Il - #e Lys Ser Cys Lys Cys          450              - #   455              - #   460                          - - Ser Ile Cys Ser Asp Ser Ile Thr His His Il - #e Tyr Glu Thr Thr Ser      465                 4 - #70                 4 - #75                 4 -      #80                                                                              - - Cys Ile Asn Tyr Lys Ser Thr Asp Asn Asp Le - #u Met Ile Val Leu        Phe                                                                                             485  - #               490  - #               495             - - Asn Leu Thr Arg Tyr Leu Met His Gly Met Il - #e His Pro Asn Leu Ile                  500      - #           505      - #           510                  - - Ser Val Lys Gly Trp Gly Pro Leu Ile Gly Le - #u Leu Thr Gly Asp Ile              515          - #       520          - #       525                      - - Gly Ile Asn Leu Lys Leu Tyr Ser Thr Met As - #n Ile Asn Gly Leu Arg          530              - #   535              - #   540                          - - Tyr Gly Asp Ile Thr Leu Ser Ser Tyr Asp Me - #t Ser Asn Lys Leu Val      545                 5 - #50                 5 - #55                 5 -      #60                                                                              - - Ser Ile Ile Asn Thr Pro Ile Tyr Glu Leu Il - #e Pro Phe Thr Thr        Cys                                                                                             565  - #               570  - #               575             - -  Cys Ser Leu Asn Glu Tyr Tyr Ser Lys Ile - #Val Ile Leu Ile Asn Val                  580      - #           585      - #           590                  - - Ile Leu Glu Tyr Met Ile Ser Ile Ile Leu Ty - #r Arg Ile Leu Ile Val              595          - #       600          - #       605                      - - Lys Arg Phe Asn Asn Ile Lys Glu Phe Ile Se - #r Lys Val Val Asn Thr          610              - #   615              - #   620                          - - Val Leu Glu Ser Ser Gly Ile Tyr Phe Cys Gl - #n Met Arg Val His Glu      625                 6 - #30                 6 - #35                 6 -      #40                                                                              - - Gln Ile Glu Leu Glu Ile Asp Glu Leu Ile Il - #e Asn Gly Ser Met        Pro                                                                                             645  - #               650  - #               655             - - Val Gln Leu Met His Leu Leu Leu Lys Val Al - #a Thr Ile Ile Leu Glu                  660      - #           665      - #           670                  - - Glu Ile Lys Glu Ile                                                              675                                                                    - -  - - (2) INFORMATION FOR SEQ ID NO:116:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 43 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Infectious - #bovine rhinotracheitis virus                      (B) STRAIN: Cooper Stra - #in                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:116:                             - - CTGGTTCGGC CCAGAATTCG ATGCAACCCA CCGCGCCGCC CCG    - #                      - # 43                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:117:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 42 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Infectious - #bovine rhinotracheitis virus                      (B) STRAIN: Cooper Stra - #in                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:117:                             - - CTCGCTCGCC CAGGATCCCT AGCGGAGGAT GGACTTGAGT CG    - #                      - #  42                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:118:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 31 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Equine In - #fluenza A neuraminidase                            (B) STRAIN: Prague/56                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:118:                             - - GGGATCCATG AATCCTAATC AAAAACTCTT T        - #                  - #              31                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:119:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 31 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Equine In - #fluenza A neuraminidase                            (B) STRAIN: Prague/56                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:119:                             - - GGGATCCTTA CGAAAAGTAT TTAATTTGTG C        - #                  - #              31                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:120:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 42 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Equine in - #fluenza A hemagglutinin                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:120:                             - - GGAGGCCTTC ATGACAGACA ACCATTATTT TGATACTACT GA    - #                      - #  42                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:121:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 40 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Equine in - #fluenza A hemagglutinin                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:121:                             - - GAAGGCCTTC TCAAATGCAA ATGTTGCATC TGATGTTGCC     - #                      - #    40                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:122:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 32 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Equine In - #fluenza A hemagglutinin                            (B) STRAIN: Prague/56                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:122:                             - - GGGATCCATG AACACTCAAA TTCTAATATT AG       - #                  - #              32                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:123:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 30 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Equine In - #fluenza A hemagglutinin                            (B) STRAIN: Prague/56                                                - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:123:                             - - GGGATCCTTA TATACAAATA GTGCACCGCA         - #                  - #               30                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:124:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 30 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Equine In - #fluenza A neuraminidase                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:124:                             - - GGGTCGACAT GAATCCAAAT CAAAAGATAA         - #                  - #               30                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:125:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 29 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Equine In - #fluenza A neuraminidase                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:125:                             - - GGGTCGACTT ACATCTTATC GATGTCAAA         - #                  - #                29                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:126:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Human                                                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:126:                             - - CTCGAATTCG AAGTGGGCAA CGTGGATCCT CGC       - #                  - #             33                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:127:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Human                                                  - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:127:                             - - CAGTTAGCCT CCCCCATCTC CCCA          - #                  - #                    24                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:128:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 21 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Equine he - #rpesvirus type 1                          - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:128:                             - - CGGAATTCCT CTGGTTGCCG T           - #                  - #                      - #21                                                                   - -  - - (2) INFORMATION FOR SEQ ID NO:129:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 22 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Equine he - #rpesvirus type 1                          - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:129:                             - - GACGGTGGAT CCGGTAGGCG GT           - #                  - #                     22                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:130:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 34 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bovine pa - #rainfluenza-3 virus                       - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:130:                             - - TTATGGATCC TGCTGCTGTG TTGAACAACT TTGT       - #                  -      #        34                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:131:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 38 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bovine pa - #rainfluenza-3 virus                       - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:131:                             - - CCGCGGATCC CATGACCATC ACAACCATAA TCATAGCC      - #                      - #     38                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:132:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 43 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bovine pa - #rainfluenza-3 virus                       - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:132:                             - - CGTCGGATCC CTTAGCTGCA GTTTTTTGGA ACTTCTGTTT TGA    - #                      - # 43                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:133:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 40 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bovine pa - #rainfluenza-3 virus                       - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:133:                             - - CATAGGATCC CATGGAATAT TGGAAACACA CAAACAGCAC     - #                      - #    40                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:134:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 42 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bovine vi - #ral diarrhea virus                                 (B) STRAIN: Singer Stra - #in                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:134:                             - - ACGTCGGATC CCTTACCAAA CCACGTCTTA CTCTTGTTTT CC    - #                      - #  42                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:135:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 40 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bovine vi - #ral diarrhea virus                                 (B) STRAIN: Singer Stra - #in                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:135:                             - - ACATAGGATC CCATGGGAGA AAACATAACA CAGTGGAACC     - #                      - #    40                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:136:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bovine vi - #ral diarrhea virus                                 (B) STRAIN: Singer Stra - #in                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:136:                             - - CGTGGATCCT CAATTACAAG AGGTATCGTC TAC       - #                  - #             33                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:137:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 31 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bovine vi - #ral diarrhea virus                                 (B) STRAIN: Singer Stra - #in                                        - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:137:                             - - CATAGATCTT GTGGTGCTGT CCGACTTCGC A        - #                  - #              31                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:138:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 37 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bovine re - #spiratory syncytial virus                          (B) STRAIN: Strain 375                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:138:                             - - TGCAGGATCC TCATTTACTA AAGGAAAGAT TGTTGAT      - #                       - #      37                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:139:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 35 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bovine re - #spiratory syncytial virus                          (B) STRAIN: Strain 375                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:139:                             - - CTCTGGATCC TACAGCCATG AGGATGATCA TCAGC       - #                       - #       35                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:140:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 40 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bovine re - #spiratory syncytial virus                          (B) STRAIN: Strain 375                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:140:                             - - CGTCGGATCC CTCACAGTTC CACATCATTG TCTTTGGGAT     - #                      - #    40                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:141:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 41 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bovine re - #spiratory syncytial virus                          (B) STRAIN: Strain 375                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:141:                             - - CTTAGGATCC CATGGCTCTT AGCAAGGTCA AACTAAATGA C    - #                      - #   41                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:142:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 41 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bovine re - #spiratory syncytial virus                          (B) STRAIN: Strain 375                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:142:                             - - CGTTGGATCC CTAGATCTGT GTAGTTGATT GATTTGTGTG A    - #                      - #   41                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:143:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 41 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (vi) ORIGINAL SOURCE:                                                          (A) ORGANISM: Bovine re - #spiratory syncytial virus                          (B) STRAIN: Strain 375                                               - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:143:                             - - CTCTGGATCC TCATACCCAT CATCTTAAAT TCAAGACATT A    - #                      - #   41                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:144:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:144:                             - - ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:145:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 128 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:145:                             - - GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TG -             #GCATATAA     60                                                                 - - ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAAATGAATT TG -            #ATCCATGA    120                                                                 - - ATCCTAAT                - #                  - #                       - #         128                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:146:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 120 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:146:                             - - CTTTTCGTAA GGATCAATTC GGATCCATAA TTAATTAATT TTTATCCCGG CG -            #CGCCTCGA     60                                                                 - - CTCTAGAATT TCATTTTGTT TTTTTCTATG CTATAAATGA ATTCGGATCC CG -            #TCGTTTTA    120                                                                 - -  - - (2) INFORMATION FOR SEQ ID NO:147:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 116 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:147:                             - - GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AA -            #AAGATCCA     60                                                                 - - TAATTAATTA ACCCGGGTCG AGGCGCGCCG GGTCGACCTG CAGGCGGCCG CT - #ATAC            116                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:148:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:148:                             - - TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:149:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:149:                             - - ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:150:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 168 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:150:                             - - GTATTGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TG -             #GCATATAA     60                                                                 - - ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAAATGAATT CA -            #CCCGCTGG    120                                                                 - - TGGCGGTCTT TGGCGCGGGC CCCGTGGGCA TCGGCCCGGG CACCACGG  - #                   168                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:151:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 112 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:151:                             - - GAGCTCGAAT TCGGATCCAT AATTAATTAA TTTTTATCCC GGCGCGCCTC GA -             #CTCTAGAA     60                                                                 - - TTTCATTTTG TTTTTTTCTA TGCTATAAAT GAATTCGGAT CCCGTCGTTT TA - #                112                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:152:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 116 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:152:                             - - GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AA -             #AAGATCCA     60                                                                 - - TAATTAATTA ACCCGGGTCG AGGCGCGCCG GGTCGACCTG CAGGCGGCCG CT - #ATAC            116                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:153:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:153:                             - - TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:154:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:154:                             - - ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:155:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 104 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:155:                             - - AAATATATAA ATACCATGTT AGAATTTGGT CTGCTGCAGG TCGACTCTAG AA -             #TTTCATTT     60                                                                 - - TGTTTTTTTC TATGCTATAA ATGAATTCGG ATCCCGTCGT TTTA   - #                      - #104                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:156:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 185 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:156:                             - - GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AA -             #AAGATCCA     60                                                                 - - TAATTAATTA ACCCGGTCGA CTCTAGAAAA AATTGAAAAA CTATTCTAAT TT -            #ATTGCACG    120                                                                 - - GAGATCTTTT TTTTTTTTTT TTTTTTGGCA TATAAATGAA TTCGGATCCC CG -            #GTGGCTTT    180                                                                 - - GGGGG                 - #                  - #                  -      #           185                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:157:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 66 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:157:                             - - CTCAATGTTA GGGTACCGAG CTCGAATTGG GTCGACCGGG TCGACCTGCA GC -            #CTACATGG     60                                                                 - - AAATCT                 - #                  - #                  -     #           66                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:158:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:158:                             - - TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:159:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:159:                             - - ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:160:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 127 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:160:                             - - GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TG -             #GCATATAA     60                                                                 - - ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAAATGAATT TC -            #GACATGAA    120                                                                 - - TCCAAAT                 - #                  - #                       - #         127                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:161:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 122 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:161:                             - - GATAAGATGT AAGTCGAAAT TCGGATCCAT AATTAATTAA TTTTTATCCC GG -            #CGCGCCTC     60                                                                 - - GACTCTAGAA TTTCATTTTG TTTTTTTCTA TGCTATAAAT GAATTCGGAT CC -            #CGTCGTTT    120                                                                 - - TA                  - #                  - #                  - #                 122                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:162:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 116 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:162:                             - - GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AA -             #AAGATCCA     60                                                                 - - TAATTAATTA ACCCGGGTCG AGGCGCGCCG GGTCGACCTG CAGGCGGCCG CT - #ATAC            116                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:163:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:163:                             - - TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:164:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:164:                             - - ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:165:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 61 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:165:                             - - GTATAGCGGC CGCCTGCAGG TCGACCTGCA GTGAATAATA AAATGTGTGT TT -             #GTCCGAAA     60                                                                 - - T                  - #                  - #                  - #                   61                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:166:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 45 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:166:                             - - CTCCATAGAA GACACCGGGA CCATGGATCC CGTCGTTTTA CAACG   - #                      - #45                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:167:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 105 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:167:                             - - TCGGCGGAAA TCCAGCTGAG CGCCGGTCGC TACCATTACC AGTTGGTCTG GT -             #GTCAAAAA     60                                                                 - - GATCTAGAAT AAGCTAGAGG ATCGATCCCC TATGGCGATC ATCAG   - #                     105                                                                        - -  - - (2) INFORMATION FOR SEQ ID NO:168:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 31 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:168:                             - - CTGCAGGTCG ACCTGCAGGC GGCCGCTATA C        - #                  - #              31                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:169:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:169:                             - - TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:170:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:170:                             - - ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:171:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 193 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:171:                             - - GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TG -             #GCATATAA     60                                                                 - - ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAAATGAATT CC -            #GAAGTGGG    120                                                                 - - CAACGTGGAT CCTCGCCCTC GGGCTCCTCG TGGTCCGCAC CGTCGTGGCC AG -            #AAGTGCTC    180                                                                 - - CTACTAGCTC GAG              - #                  - #                      - #     193                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:172:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 123 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:172:                             - - ATCATTAGCA CGTTAACTTA ATAAGATCCA TAATTAATTA ATTTTTATCC CG -             #GCGCGCCT     60                                                                 - - CGACTCTAGA ATTTCATTTT GTTTTTTTCT ATGCTATAAA TGAATTCGGA TC -            #CCGTCGTT    120                                                                 - - TTA                  - #                  - #                  - #                123                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:173:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 116 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:173:                             - - GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AA -             #AAGATCCA     60                                                                 - - TAATTAATTA ACCCGGGTCG AGGCGCGCCG GGTCGACCTG CAGGCGGCCG CT - #ATAC            116                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:174:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:174:                             - - TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:175:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:175:                             - - ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:176:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 133 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:176:                             - - GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TG -             #GCATATAA     60                                                                 - - ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAAATGAATT CC -            #TCTGGTTG    120                                                                 - - CCGTTCTGTC GGC              - #                  - #                      - #     133                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:177:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 99 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:177:                             - - GAAAATGAAA AAATGGTTTA AACCGGGGGC GCGCCTCGAC TCTAGAATTT CA -             #TTTTGTTT     60                                                                 - - TTTTCTATGC TATAAATGAA TTCGGATCCC GTCGTTTTA      - #                      - #    99                                                                     - -  - - (2) INFORMATION FOR SEQ ID NO:178:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 116 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:178:                             - - GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AA -             #AAGATCCA     60                                                                 - - TAATTAATTA ACCCGGGTCG AGGCGCGCCG GGTCGACCTG CAGGCGGCCG CT - #ATAC            116                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:179:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:179:                             - - TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:180:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:180:                             - - ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:181:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 140 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:181:                             - - GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TG -             #GCATATAA     60                                                                 - - ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAAATGAATT CG -            #GATCAGCT    120                                                                 - - TATGATGGAT GGACGTTTGG            - #                  - #                      - #140                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:182:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 123 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:182:                             - - GGAGGTGTCC ACGGCCTTAA AGCTGATCCA TAATTAATTA ATTTTTATCC CG -             #GCGCGCCT     60                                                                 - - CGACTCTAGA ATTTCATTTT GTTTTTTTCT ATGCTATAAA TGAATTCGGA TC -            #CCGTCGTT    120                                                                 - - TTA                  - #                  - #                  - #                123                                                                  - -  - - (2) INFORMATION FOR SEQ ID NO:183:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 116 base - #pairs                                                 (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:183:                             - - GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AA -             #AAGATCCA     60                                                                 - - TAATTAATTA ACCCGGGTCG AGGCGCGCCG GGTCGACCTG CAGGCGGCCG CT - #ATAC            116                                                                       - -  - - (2) INFORMATION FOR SEQ ID NO:184:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 51 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:184:                             - - TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C - #                 51                                                                         - -  - - (2) INFORMATION FOR SEQ ID NO:185:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 39 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:185:                             - - GAAGCATGCC CGTTCTTATC AATAGTTTAG TCGAAAATA      - #                      - #    39                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:186:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 41 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:186:                             - - CATAAGATCT GGCATTGTGT TATTATACTA ACAAAAATAA G    - #                      - #   41                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:187:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 41 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:187:                             - - CCGTAGTCGA CAAAGATCGA CTTATTAATA TGTATGGGAT T    - #                      - #   41                                                                      - -  - - (2) INFORMATION FOR SEQ ID NO:188:                                   - -      (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 39 base - #pairs                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: double                                                      (D) TOPOLOGY: linear                                                 - -     (ii) MOLECULE TYPE: DNA (genomic)                                     - -    (iii) HYPOTHETICAL: NO                                                 - -     (iv) ANTI-SENSE: NO                                                   - -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:188:                             - - GCCTGAAGCT TCTAGTACAG TATTTACGAC TTTTGAAAT      - #                      - #    39                                                                    __________________________________________________________________________

What is claimed is:
 1. A recombinant swinepox virus which comprises aforeign DNA which (a) is inserted into a non-essential site located inthe HindIII M fragment of the swinepox genome, wherein the foreign DNAencodes a polypeptide derived from a human pathogen and (b) is expressedin a host cell infected by the recombinant swinepox virus.
 2. Therecombinant swinepox virus of claim 1, wherein the polypeptide isderived from the group consisting of human herpesvirus, herpes simplexvirus-1, herpes simplex virus-2, human cytomegalovirus, Epstein-Barrvirus, Varicell-Zoster virus, human herpesvirus-6, human herpesvirus- 7,human influenza, human immunodeficiency virus, rabies virus, measlesvirus, hepatitis B virus and hepatitis C virus.
 3. The recombinantswinepox virus of claim 2, wherein the polypeptide is hepatitis B viruscore protein or hepatitis B virus surface protein.
 4. The recombinantswinepox virus of claim 3, designated S-SPV-031.
 5. The recombinantswinepox virus of claim 1, wherein the polypeptide is associated withmalaria.
 6. The recombinant swinepox virus of claim 1, wherein theinsertion site is present within the larger HindIII to BglIIsub-fragment of the HindIII M fragment of the swinepox viral genome. 7.The recombinant swinepox virus of claim 6, wherein the insertion site iswithin an open reading frame contained in the HindIII to BglIIsub-fragment.
 8. The recombinant swinepox virus of claim 7, wherein theinsertion site is the AccI restriction endonuclease site located in theHindIII to BglII sub-fragment.
 9. The recombinant swinepox virus ofclaim 8, wherein the AccI restriction endonuclease site is replaced by aNotI restriction endonuclease site.
 10. The recombinant swinepox virusof claim 8, wherein the AccI restriction endonuclease site is replacedby a PstI restriction endonuclease site.
 11. The recombinant swinepoxvirus of claim 1, further comprising an insertion site within an openreading frame encoding swinepox virus thymidine kinase.
 12. Therecombinant swinepox virus of claim 11, wherein the insertion site isthe NdeI restriction endonuclease site located within the open readingframe encoding the swinepox virus thymidine kinase.
 13. The recombinantswinepox virus of claim 12, wherein the NdeI restriction site isreplaced by a AscI restriction endonuclease site.
 14. The recombinantswinepox virus of claim 1, wherein the expression of the foreign DNA isunder the control of a promoter located upstream from the foreign DNA.15. The recombinant swinepox virus of claim 14, wherein the promoter isan endogenous swinepox viral promoter or an exogenous promoter.
 16. Therecombinant swinepox virus of claim 15, wherein the exogenous promoteris a synthetic pox viral promoter.
 17. The recombinant swinepox virus ofclaim 15, wherein the exogenous promoter is human cytomegalovirusimmediately early gene promoter.
 18. A recombinant swinepox virus whichcomprises a foreign DNA which (a) is inserted into a non-essential sitelocated in the HindIII M fragment of the swinepox genome, wherein theforeign DNA encodes a cytokine capable of stimulating an immune responsein a host infected by the recombinant swinepox virus and (b) isexpressed in a host cell infected by the recombinant swinepox virus. 19.The recombinant swinepox virus of claim 18, wherein the cytokine isinterleukin-2, interleukin-6, interleukin-12, an interferon, agranulocyte-macrophage colony stimulating factor, or an interleukinreceptor.
 20. The recombinant swinepox virus of claim 19, wherein thecytokine is human interleukin-2.
 21. A recombinant swinepox virus whichcomprises a foreign DNA which (a) is inserted into a non-essential siteof the swinepox genome, wherein the foreign DNA encodes a polypeptidederived from an equine pathogen and (b) is expressed in a host cellinfected by the recombinant swinepox virus.
 22. The recombinant swinepoxvirus of claim 21, wherein the polypeptide is derived from equineinfluenza virus or equine herpesvirus.
 23. The recombinant swinepoxvirus of claim 22, wherein the polypeptide is equine influenza virustype A/Alaska 91 neuraminidase, equine influenza virus type A/Prague 56neuraminidase, equine influenza virus type A/Miami 63 neuraminidase,equine influenza virus type A/Kentucky neuraminidase, equine herpesvirustype 1 glycoprotein B, or equine herpesvirus type 1 glycoprotein D. 24.The recombinant swinepox virus of claim 23, wherein the polypeptide isequine influenza virus type A/Alaska 91 neuraminidase.
 25. Therecombinant swinepox virus of claim 24, designated S-SPV-033.
 26. Therecombinant swinepox virus of claim 23, wherein the polypeptide isequine influenza virus type A/Prague 56 neuraminidase.
 27. Therecombinant swinepox virus of claim 26, designated S-SPV-034.
 28. Therecombinant swinepox virus of claim 23, wherein the polypeptide isequine herpesvirus type 1 glycoprotein B.
 29. The recombinant swinepoxvirus of claim 28, designated S-SPV-038.
 30. The recombinant swinepoxvirus of claim 23, wherein the polypeptide is equine herpesvirus type 1glycoprotein D.
 31. The recombinant swinepox virus of claim 30,designated S-SPV-039.
 32. A recombinant swinepox virus which comprises aforeign DNA which (a) is inserted into a non-essential site located inthe HindIII M fragment of the swinepox genome, wherein the foreign DNAencodes a polypeptide derived from bovine respiratory syncytial virus orbovine parainfluenza virus, and wherein the foreign DNA (b) is expressedin a host cell infected by the recombinant swinepox virus.
 33. Therecombinant swinepox virus of claim 32, wherein the polypeptide isbovine respiratory syncytial virus attachment protein (BRSV G), bovinerespiratory syncytial virus fusion protein (BRSV F), bovine respiratorysyncytial virus nucleocapsid protein (BRSV N), bovine parainfluenzavirus type 3 fusion protein, or bovine parainfluenza virus type 3hemagglutinin neuraminidase.
 34. The recombinant swinepox virus of claim33, wherein the polypeptide is bovine respiratory syncytial virusattachment protein (BRSV G).
 35. The recombinant swinepox virus of claim34, designated S-SPV-020.
 36. The recombinant swinepox virus of claim33, wherein the polypeptide is bovine respiratory syncytial virus fusionprotein (BRSV F).
 37. The recombinant swinepox virus of claim 36,designated S-SPV-029.
 38. The recombinant swinepox virus of claim 33,wherein the polypeptide is bovine respiratory syncytial virusnucleocapsid protein (BRSV N).
 39. The recombinant swinepox virus ofclaim 38, designated S-SPV-030.
 40. The recombinant swinepox virus ofclaim 33, wherein the polypeptide is bovine parainfluenza virus type 3fusion protein.
 41. The recombinant swinepox virus of claim 40,designated S-SPV-028.
 42. The recombinant swinepox virus of claim 32,wherein the polypeptide is bovine parainfluenza virus type 3hemagglutinin neuraminidase.
 43. A recombinant swinepox virus whichcomprises a foreign DNA which (a) is inserted into a non-essential sitelocated in the HindIII M fragment of the swinepox genome wherein theforeign DNA encodes bovine viral diarrhea virus glycoprotein 48 orbovine viral diarrhea virus glycoprotein 53, and wherein the foreign DNA(b) is expressed in a host infected by the recombinant swinepox virus.44. The recombinant swinepox virus of claim 43, designated S-SPV-032.45. The recombinant swinepox virus of claim 43, designated S-SPV-040.46. A recombinant swinepox virus which comprises a foreign DNA which (a)is inserted into a non-essential site located in the HindIII M fragmentof the swinepox genome, wherein the foreign DNA encodes a polypeptidederived from infectious bursal disease virus and wherein the foreign DNA(b) is expressed in a host cell infected by the recombinant swinepoxvirus.
 47. The recombinant swinepox virus of claim 46, wherein thepolypeptide is infectious bursal disease virus polyprotein.
 48. Therecombinant swinepox virus of claim 46, wherein the polypeptide isinfectious bursal disease virus VP2.